Integrated top-down and bottom-up Proteomic Approach to Characterize The Antigen-Binding

An integrated top-down and bottom-up proteomic approach to characterize the antigen-binding fragment of antibodies.

We have previously shown that different individuals exposed to the same antigen produce antibodies with identical mutations in their complementarity determining regions (CDR), suggesting that CDR tryptic peptides can serve as biomarkers for disease diagnosis and prognosis.

Complete Fabs derived from disease-specific antibodies have even higher potential; they could potentially be used for disease treatment and are required to identify the antigens toward which the antibodies are directed. However, complete Fab sequence characterization via LC-MS analysis of tryptic peptides (i.e. bottom-up) has proven to be impractical for mixtures of antibodies.

To tackle this challenge, we have developed an integrated bottom-up and top-down MS approach, employing 2D chromatography coupled with Fourier transform mass spectrometry (FTMS), and applied this approach for full characterization of the variable parts of two pharmaceutical monoclonal antibodies with sensitivity comparable to the bottom-up standard.

These efforts represent an essential step toward the identification of disease specific antibodies in patient samples with potentially significant clinical impact.

Selective 351 nm photodissociation of cysteine-containing peptides for discrimination of antigen-binding regions of IgG fragments in bottom-up liquid chromatography-tandem mass spectrometry workflows.

Despite tremendous inroads in the development of more sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) strategies for mass spectrometry-based proteomics, there remains a significant need for enhancing the selectivity of MS/MS-based workflows for streamlined analysis of complex biological mixtures.

Here, a novel LC-MS/MS platform based on 351 nm ultraviolet photodissociation (UVPD) is presented for the selective analysis of cysteine-peptide subsets in complex protein digests. Cysteine-selective UVPD is mediated through the site-specific conjugation of reduced cysteine residues with a 351 nm active chromogenic Alexa Fluor 350 (AF350) maleimide tag.

Only peptides containing the AF350 chromophore undergo photodissociation into extensive arrays of b- and y-type fragment ions, thus providing a facile means for differentiating cysteine-peptide targets from convoluting peptide backgrounds.

With the use of this approach in addition to strategic proteolysis, the selective analysis of diagnostic heavy-chain complementarity determining regions (CDRs) of single-chain antibody (scAb) fragments is demonstrated.

Fragments of the Roelofs effect: a bottom-up effect equal to the sum of its parts.

The Roelofs effect is a distortion of perceived space that occurs when a large frame whose center is offset left or right from the objective midline is presented visually to an observer and causes a bias in the observer’s subjective judgment of midline.

Experiments were designed to test whether an isolated fragment (left or right end) of a Roelofs-inducing frame was capable of generating the Roelofs effect and to determine whether prior experience with intact frames would provide a top-down influence that would bias the Roelofs effect resulting from fragment presentation.

Although the fragments did induce an effect, top-down information did not play a significant role even after a 5-day training paradigm. Instead, we found that the effect generated by an intact frame was equal in magnitude to the sum of the effects generated by the individual fragments.

In addition, perception was found to be differentially affected by the two ends of the frame, with fragments falling in the right visual field causing a larger effect than those falling in the left.

Goat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E06R0362-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E06R0362-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E08R0362-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E08R0362-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E08R0362-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E07R0362-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E07R0362-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E07R0362-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E03R0362-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E03R0362-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E03R0362-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E02R0362-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Rat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E02R0362-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Rat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E02R0362-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Rat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E09R0362-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E09R0362-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E09R0362-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Native Aspergillus oryzae endo-Inulinase (food grade)
NATE-1246	Creative Enzymes	1kg	EUR 324

Native Aspergillus oryzae exo-Inulinase (Food Grade)
NATE-1250	Creative Enzymes	1kg	EUR 324

Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E05R0362-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E05R0362-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E05R0362-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Recombinant Aspergillus Oryzae EFG1 Protein (aa 1-320) [His]
VAng-Wyb3967-1mg	Creative Biolabs	1 mg	EUR 5378.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Efg1p, recombinant protein.

Recombinant Aspergillus Oryzae Egd2 Protein (aa 1-202) [His]
VAng-Wyb3968-1mg	Creative Biolabs	1 mg	EUR 4820.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Nascent Polypeptide-Associated Complex Subunit alpha, recombinant protein.

Recombinant Aspergillus Oryzae EIF3E Protein (aa 1-451) [His]
VAng-Wyb3969-1mg	Creative Biolabs	1 mg	EUR 5998.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Eukaryotic Translation Initiation Factor 3 Subunit E, recombinant protein.

Recombinant Aspergillus Oryzae EIF3G Protein (aa 1-287) [His]
VAng-Wyb3970-1mg	Creative Biolabs	1 mg	EUR 5220
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Eukaryotic Translation Initiation Factor 3, Subunit G, recombinant protein.

Recombinant Aspergillus Oryzae EIF3I Protein (aa 1-335) [His]
VAng-Wyb3971-1mg	Creative Biolabs	1 mg	EUR 5448
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Eukaryotic Translation Initiation Factor 3, Subunit I, recombinant protein.

Recombinant Aspergillus Oryzae ESF2 Protein (aa 1-335) [His]
VAng-Wyb3972-1mg	Creative Biolabs	1 mg	EUR 5448
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Esf2p, recombinant protein.

Recombinant Aspergillus Oryzae FIS1 Protein (aa 1-126) [His]
VAng-Wyb3973-1mg	Creative Biolabs	1 mg	EUR 4459.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fis1p, recombinant protein.

Recombinant Aspergillus Oryzae FPR1 Protein (aa 1-116) [His]
VAng-Wyb3974-1mg	Creative Biolabs	1 mg	EUR 4412.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fpr1p, recombinant protein.

Recombinant Aspergillus Oryzae FPR2 Protein (aa 20-134) [His]
VAng-Wyb3975-1mg	Creative Biolabs	1 mg	EUR 4407.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fpr2p, recombinant protein.

Recombinant Aspergillus Oryzae FPR4 Protein (aa 1-470) [His]
VAng-Wyb3976-1mg	Creative Biolabs	1 mg	EUR 6091.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fpr4p, recombinant protein.

Recombinant Aspergillus Oryzae GAPDH Protein (aa 1-338) [His]
VAng-Wyb3977-1mg	Creative Biolabs	1 mg	EUR 5463.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Glyceraldehyde-3-Phosphate Dehydrogenase, recombinant protein.

Recombinant Aspergillus Oryzae GCD14 Protein (aa 1-474) [His]
VAng-Wyb3978-1mg	Creative Biolabs	1 mg	EUR 6108
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Gcd14p, recombinant protein.

Recombinant Aspergillus Oryzae GINS2 Protein (aa 1-269) [His]
VAng-Wyb3979-1mg	Creative Biolabs	1 mg	EUR 5133.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) GINS Complex Subunit 2 (Psf2 Homolog), recombinant protein.

Recombinant Aspergillus Oryzae H2A Protein (aa 2-133) [His]
VAng-Wyb3980-1mg	Creative Biolabs	1 mg	EUR 4488
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Histone H2A, recombinant protein.

Recombinant Aspergillus Oryzae HCR1 Protein (aa 1-265) [His]
VAng-Wyb3981-1mg	Creative Biolabs	1 mg	EUR 5119.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) COenzyme Q (Ubiquinone) Hcr1p, recombinant protein.

Recombinant Aspergillus Oryzae JIP5 Protein (aa 1-417) [His]
VAng-Wyb3982-1mg	Creative Biolabs	1 mg	EUR 5840.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Jip5p, recombinant protein.

Recombinant Aspergillus Oryzae KAE1 Protein (aa 1-358) [His]
VAng-Wyb3983-1mg	Creative Biolabs	1 mg	EUR 5563.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Kae1p, recombinant protein.

Recombinant Aspergillus Oryzae LCL2 Protein (aa 22-1226) [His]
VAng-Wyb3984-1mg	Creative Biolabs	1 mg	EUR 4339.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Lcl2p, recombinant protein.

Recombinant Aspergillus Oryzae LIA1 Protein (aa 1-339) [His]
VAng-Wyb3985-1mg	Creative Biolabs	1 mg	EUR 5470.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Lia1p, recombinant protein.

Recombinant Aspergillus Oryzae LOC1 Protein (aa 1-190) [His]
VAng-Wyb3986-1mg	Creative Biolabs	1 mg	EUR 4765.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) COenzyme Q (Ubiquinone) Loc1p, recombinant protein.

Recombinant Aspergillus Oryzae LOC100799412 Protein (aa 1-344) [His]
VAng-Wyb3987-1mg	Creative Biolabs	1 mg	EUR 5493.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mitogen-Activated Protein Kinase MpkC, recombinant protein.

Recombinant Aspergillus Oryzae MDE1 Protein (aa 1-241) [His]
VAng-Wyb3988-1mg	Creative Biolabs	1 mg	EUR 5006.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mde1p, recombinant protein.

Recombinant Aspergillus Oryzae MED21 Protein (aa 1-208) [His]
VAng-Wyb3989-1mg	Creative Biolabs	1 mg	EUR 4850.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 21, recombinant protein.

Recombinant Aspergillus Oryzae MED6 Protein (aa 1-328) [His]
VAng-Wyb3990-1mg	Creative Biolabs	1 mg	EUR 5421.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 6, recombinant protein.

Recombinant Aspergillus Oryzae MED7 Protein (aa 1-261) [His]
VAng-Wyb3991-1mg	Creative Biolabs	1 mg	EUR 5101.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 7, recombinant protein.

Recombinant Aspergillus Oryzae MED8 Protein (aa 1-244) [His]
VAng-Wyb3992-1mg	Creative Biolabs	1 mg	EUR 5022
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 8, recombinant protein.

Recombinant Aspergillus Oryzae METTL1 Protein (aa 1-330) [His]
VAng-Wyb3993-1mg	Creative Biolabs	1 mg	EUR 5425.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Methyltransferase Like 1, recombinant protein.

Recombinant Aspergillus Oryzae MMM1 Protein (aa 47-494) [His]
VAng-Wyb3994-1mg	Creative Biolabs	1 mg	EUR 5989.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mmm1p, recombinant protein.

Recombinant Aspergillus Oryzae MOCS2 Protein (aa 1-179) [His]
VAng-Wyb3995-1mg	Creative Biolabs	1 mg	EUR 4708.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Molybdenum Cofactor Synthesis 2, recombinant protein.

Recombinant Aspergillus Oryzae Mri1 Protein (aa 1-381) [His]
VAng-Wyb3996-1mg	Creative Biolabs	1 mg	EUR 5668.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Methylthioribose-1-Phosphate Isomerase Homolog, recombinant protein.

Recombinant Aspergillus Oryzae MZM1 Protein (aa 19-114) [His]
VAng-Wyb3997-1mg	Creative Biolabs	1 mg	EUR 4316.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mzm1p, recombinant protein.

Recombinant Aspergillus Oryzae NBP35 Protein (aa 1-325) [His]
VAng-Wyb3998-1mg	Creative Biolabs	1 mg	EUR 5402.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Nbp35p, recombinant protein.

Recombinant Aspergillus Oryzae NGLY1 Protein (aa 1-457) [His]
VAng-Wyb3999-1mg	Creative Biolabs	1 mg	EUR 6028.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) N-Glycanase 1, recombinant protein.

Recombinant Aspergillus Oryzae NUCS Protein (aa 21-287) [His]
VAng-Wyb4000-1mg	Creative Biolabs	1 mg	EUR 5131.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) NucS Gene Product, recombinant protein.

Recombinant Aspergillus Oryzae NUF2 Protein (aa 1-463) [His]
VAng-Wyb4001-1mg	Creative Biolabs	1 mg	EUR 6057.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Spindle Pole Body Protein, recombinant protein.

Recombinant Aspergillus Oryzae P4HB Protein (aa 21-515) [His]
VAng-Wyb4002-1mg	Creative Biolabs	1 mg	EUR 6210
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Prolyl 4-Hydroxylase, beta Polypeptide, recombinant protein.

Recombinant Aspergillus Oryzae PNO1 Protein (aa 1-260) [His]
VAng-Wyb4003-1mg	Creative Biolabs	1 mg	EUR 5095.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Pno1p, recombinant protein.

Recombinant Aspergillus Oryzae POA1 Protein (aa 1-201) [His]
VAng-Wyb4004-1mg	Creative Biolabs	1 mg	EUR 4818
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Poa1p, recombinant protein.

Recombinant Aspergillus Oryzae PPME1 Protein (aa 1-427) [His]
VAng-Wyb4005-1mg	Creative Biolabs	1 mg	EUR 5887.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Protein Phosphatase Methylesterase 1, recombinant protein.

Recombinant Aspergillus Oryzae PXR1 Protein (aa 1-306) [His]
VAng-Wyb4006-1mg	Creative Biolabs	1 mg	EUR 5312.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Pxr1p, recombinant protein.

Recombinant Aspergillus Oryzae RBBP7 Protein (aa 1-436) [His]
VAng-Wyb4007-1mg	Creative Biolabs	1 mg	EUR 5932.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Retinoblastoma Binding Protein 7, recombinant protein.

Recombinant Aspergillus Oryzae RMT2 Protein (aa 1-413) [His]
VAng-Wyb4008-1mg	Creative Biolabs	1 mg	EUR 5817.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rmt2p, recombinant protein.

Recombinant Aspergillus Oryzae RPS1 Protein (aa 2-256) [His]
VAng-Wyb4009-1mg	Creative Biolabs	1 mg	EUR 5072.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rps1 Gene Product, recombinant protein.

Recombinant Aspergillus Oryzae RRD1 Protein (aa 1-478) [His]
VAng-Wyb4010-1mg	Creative Biolabs	1 mg	EUR 6130.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrd1p, recombinant protein.

Recombinant Aspergillus Oryzae RRD2 Protein (aa 1-422) [His]
VAng-Wyb4011-1mg	Creative Biolabs	1 mg	EUR 5864.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrd2p, recombinant protein.

Recombinant Aspergillus Oryzae RRG9 Protein (aa 54-284) [His]
VAng-Wyb4012-1mg	Creative Biolabs	1 mg	EUR 4959.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrg9p, recombinant protein.

Recombinant Aspergillus Oryzae RRP36 Protein (aa 1-352) [His]
VAng-Wyb4013-1mg	Creative Biolabs	1 mg	EUR 5534.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrp36p, recombinant protein.

Recombinant Aspergillus Oryzae SFH5 Protein (aa 1-455) [His]
VAng-Wyb4014-1mg	Creative Biolabs	1 mg	EUR 6018
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Sfh5p, recombinant protein.

Recombinant Aspergillus Oryzae Slx1 Protein (aa 1-406) [His]
VAng-Wyb4015-1mg	Creative Biolabs	1 mg	EUR 5791.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Structure-Specific Endonuclease Subunit Slx1, recombinant protein.

Recombinant Aspergillus Oryzae SNX3 Protein (aa 1-142) [His]
VAng-Wyb4016-1mg	Creative Biolabs	1 mg	EUR 4537.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Sorting Nexin 3, recombinant protein.

Recombinant Aspergillus Oryzae SOD1 Protein (aa 2-154) [His]
VAng-Wyb4017-1mg	Creative Biolabs	1 mg	EUR 4591.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Triosephosphate Isomerase 1, recombinant protein.

Recombinant Aspergillus Oryzae srb5 Protein (aa 1-273) [His]
VAng-Wyb4018-1mg	Creative Biolabs	1 mg	EUR 5157.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator of RNA polymerase II transcription subunit 18, recombinant protein.

Recombinant Aspergillus Oryzae SSN8 Protein (aa 1-301) [His]
VAng-Wyb4019-1mg	Creative Biolabs	1 mg	EUR 5289.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) COenzyme Q (Ubiquinone) Ssn8p, recombinant protein.

Recombinant Aspergillus Oryzae SSU72 Protein (aa 1-262) [His]
VAng-Wyb4020-1mg	Creative Biolabs	1 mg	EUR 5108.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) SSU72 RNA Polymerase II CTD Phosphatase Homolog, recombinant protein.

Recombinant Aspergillus Oryzae STS1 Protein (aa 1-312) [His]
VAng-Wyb4021-1mg	Creative Biolabs	1 mg	EUR 5338.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Sts1p, recombinant protein.

Recombinant Aspergillus Oryzae TEF1 Protein (aa 1-460) [His]
VAng-Wyb4022-1mg	Creative Biolabs	1 mg	EUR 6045.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Tef1p, recombinant protein.

Recombinant Aspergillus Oryzae TMA22 Protein (aa 1-194) [His]
VAng-Wyb4023-1mg	Creative Biolabs	1 mg	EUR 4782
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Tma22p, recombinant protein.

Recombinant Aspergillus Oryzae TPI1 Protein (aa 1-251) [His]
VAng-Wyb4024-1mg	Creative Biolabs	1 mg	EUR 5048.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Triosephosphate Isomerase 1, recombinant protein.

Recombinant Aspergillus Oryzae TRM82 Protein (aa 1-493) [His]
VAng-Wyb4025-1mg	Creative Biolabs	1 mg	EUR 6199.2
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Trm82p, recombinant protein.

Recombinant Aspergillus Oryzae UBA4 Protein (aa 1-463) [His]
VAng-Wyb4026-1mg	Creative Biolabs	1 mg	EUR 6057.6
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) COenzyme Q (Ubiquinone) Uba4p, recombinant protein.

Recombinant Aspergillus Oryzae Urm1 Protein (aa 1-120) [His]
VAng-Wyb4027-1mg	Creative Biolabs	1 mg	EUR 4428
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Urm1, recombinant protein.

Recombinant Aspergillus Oryzae XLNA Protein (aa 20-232) [His]
VAng-Wyb4028-1mg	Creative Biolabs	1 mg	EUR 4874.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) XlnA Gene Product, recombinant protein.

Recombinant Aspergillus Oryzae XlnB Protein (aa 20-221) [His]
VAng-Wyb4029-1mg	Creative Biolabs	1 mg	EUR 4820.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Probable Endo-1,4-beta-Xylanase B, recombinant protein.

Recombinant Aspergillus Oryzae ACT1 Protein (aa 1-375) [His]
VAng-Wyb3947-1mg	Creative Biolabs	1 mg	EUR 5642.4
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Actin, recombinant protein.

Recombinant Aspergillus Oryzae ADH5 Protein (aa 1-383) [His]
VAng-Wyb3948-1mg	Creative Biolabs	1 mg	EUR 5676
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Alcohol Dehydrogenase 5 (Class III), chi Polypeptide, recombinant protein.

Recombinant Aspergillus Oryzae Amy3 Protein (aa 22-499) [His]
VAng-Wyb3949-1mg	Creative Biolabs	1 mg	EUR 6130.8
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) alpha-Amylase A Type-3, recombinant protein.

Induction of cell cycle arrest and apoptosis by tomentosin in hepatocellular carcinoma HepG2 and Huh7 cells.

Tomentosin, a sesquiterpene lactone, is known to possess various biological activities. However, its anticarcinogenic activity against human hepatocellular carcinoma (HCC) cells has not been investigated in detail. Thus, this study aimed to elucidate the cytotoxic mechanism of tomentosin in human HCC cell lines HepG2 and Huh7.

WST-1, cell counting, and colony formation assay results showed that treatment with tomentosin decreased the viability and suppressed the proliferation rate of HepG2 and Huh7 cells in a dose- and time-dependent manner. Cell cycle analysis revealed increased population of cells at the SubG1 and G2/M stage, and decreased population of cells at the G0/1 stage in HepG2 and Huh7 cells treated with tomentosin.

Annexin V/propidium iodide double staining and TUNEL assay results showed increased apoptotic cell population and DNA fragmentation in HepG2 and Huh7 cells treated with tomentosin.

Western blotting analysis results showed that tomentosin treatment significantly increased the expression level of Bax, Bim (short form), cleaved PARP1, FOXO3, p53, pSer15p53, pSer20p53, pSer46p53, p21, and p27, but decreased the expression of Bcl2, caspase3, caspase7, caspase9, cyclin-dependent kinase 2 (CDK2), CDK4, CDK6, cyclinB1, cyclinD1, cyclinD2, cyclinD3, and cyclinE in a dose-dependent manner.

Taken together, this study revealed that tomentosin, which acted through cell cycle arrest and apoptosis, may be a useful therapeutic option against HCC.

Keywords: FOXO3; Tomentosin; apoptosis; cell cycle arrest; hepatocellular carcinoma; p53.