Product Info Summary
SKU: | M00227-1 |
---|---|
Size: | 100 μl |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-GAPDH Rabbit Monoclonal Antibody
SKU/Catalog Number
M00227-1
Size
100 μl
Form
Liquid
Description
Boster Bio Anti-GAPDH Rabbit Monoclonal Antibody catalog # M00227-1. Tested in WB, IHC, ICC/IF, Flow Cytometry applications. This antibody reacts with Human, Monkey, Mouse, Rat.
Storage & Handling
Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-GAPDH Rabbit Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00227-1)
Host
Rabbit
Contents
Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clonality
Monoclonal
Clone Number
BG-7
Isotype
Rabbit IgG
Immunogen
A synthesized peptide derived from human GAPDH
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
M00227-1 is reactive to GAPDH in Human, Mouse, Rat
Reconstitution
Restore with deionized water (or equivalent) for reconstitution volume of 1.0 mL
Observed Molecular Weight
36 kDa
Calculated molecular weight
36053 MW
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M00227-1 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
WB 1:3000-1:10000
IHC 1:100-1:250
ICC/IF 1:100-1:250
FC 1:50
Positive Control
WB: human Hela whole cell, human K562 whole cell, human MCF-7 whole cell, human A549 whole cell, human PC-3 whole cell, monkey COS-7 whole cell, rat liver tissue, rat brain tissue, rat RH35 whole cell, mouse liver tissue, mouse brain tissue, mouse NIH/3T3 whole cell
IHC: human bladder cancer tissue
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of GAPDH using anti-GAPDH antibody (M00227-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human PC-3 whole cell lysates,
Lane 6: monkey COS-7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH antigen affinity purified monoclonal antibody (Catalog # M00227-1) at 1:3000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GAPDH at approximately 36 kDa. The expected band size for GAPDH is at 36 kDa.
Click image to see more details
Figure 2. Western blot analysis of GAPDH using anti-GAPDH antibody (M00227-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat RH35 whole cell lysates,
Lane 4: mouse liver tissue lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH antigen affinity purified monoclonal antibody (Catalog # M00227-1) at 1:3000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GAPDH at approximately 36 kDa. The expected band size for GAPDH is at 36 kDa.
Click image to see more details
Immunohistochemical analysis of paraffin-embedded human bladder cancer, using GAPDH Antibody.
Click image to see more details
Immunofluorescent analysis of Hela cells, using GAPDH Antibody.
Click image to see more details
Immunofluorescent analysis using the Antibody at 1:50 dilution.
Click image to see more details
Immunofluorescent analysis using the Antibody at 1:50 dilution.
Protein Target Info & Infographic
Gene/Protein Information For GAPDH (Source: Uniprot.org, NCBI)
Gene Name
GAPDH
Full Name
Glyceraldehyde-3-phosphate dehydrogenase
Weight
36053 MW
Superfamily
glyceraldehyde-3-phosphate dehydrogenase family
Alternative Names
Glyceraldehyde-3-phosphate dehydrogenase;GAPDH;1.2.1.12;Peptidyl-cysteine S-nitrosylase GAPDH;2.6.99.-;GAPDH;GAPD;CDABP0047, OK/SW-cl.12; GAPDH G3PD, GAPD, HEL-S-162eP glyceraldehyde-3-phosphate dehydrogenase glyceraldehyde-3-phosphate dehydrogenase|OCAS, p38 component|Oct1 coactivator in S phase, 38 Kd component|aging-associated gene 9 protein|epididymis secretory sperm binding protein Li 162eP|peptidyl-cysteine S-nitrosylase GAPDH
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on GAPDH, check out the GAPDH Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for GAPDH: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-GAPDH Rabbit Monoclonal Antibody (M00227-1)
Hello CJ!
M00227-1 has been cited in 150 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Wang J,Fang Y,Liu YF,Wang X,Wang XL,Wang RY,Meng ZD.MiR-154 inhibits cells proliferation and metastasis in melanoma by targeting AURKA and serves as a novel prognostic indicator.Eur Rev Med Pharmacol Sci.2019 May;23(10):4275-4284.doi:10.26355/eurrev_201905_17932.PMID:31173299.
Species: Human
M00227-1 usage in article: APP:WB, SAMPLE:A375 CELL, DILUTION:1:1500
Duan Y,Tan Z,Yang M,Li J,Liu C,Wang C,Zhang F,Jin Y,Wang Y,Zhu L.PC-3-Derived Exosomes Inhibit Osteoclast Differentiation by Downregulating miR-214 and Blocking NF-κB Signaling Pathway.Biomed Res Int.2019 Apr 1;2019:8650846.doi:10.1155/2019/8650846. PMID:31058194;PMCID:PMC6463683.
Species: Human
M00227-1 usage in article: APP:WB, SAMPLE:OSTEOCLASTS, DILUTION:NA
Huang W,Guo L,Zhao M,Zhang D,Xu H,Nie Q.The Inhibition on MDFIC and PI3K/AKT Pathway Caused by miR-146b-3p Triggers Suppression of Myoblast Proliferation and Differentiation and Promotion of Apoptosis.Cells.2019 Jun 29;8(7):656.doi:10.3390/cells8070656.PMID:31261950; PMCID:PMC6678156.
Species: Chicken
M00227-1 usage in article: APP:WB, SAMPLE:QM-7 CELL, DILUTION:1:2000
Xiang W,Jiang T,Hao X, Wang R,Yao X,Sun K,Guo F,Xu T.Primary cilia and autophagy interaction is involved in mechanical stress mediated cartilage development via ERK/mTOR axis.Life Sci.2019 Feb 1;218:308-313.doi:10.1016/j. lfs.2019.01.001.Epub 2019 Jan 3.PMID:30610869.
Species: Rat
M00227-1 usage in article: APP:WB, SAMPLE:CHONDROCYTES, DILUTION:1:400
Sheng X,Li Y,Li Y,Liu W,Lu Z,Zhan J,Xu M,Chen L,Luo X,Cai G,Zhang S.PLOD2 contributes to drug resistance in laryngeal cancer by promoting cancer stem cell-like characteristics.BMC Cancer.2019 Aug 27;19(1):840.doi:10.1186/s12885-019-6029-y.PMID:31455288; PMCID:PMC6712771.
Species: Human,Mouse
M00227-1 usage in article: APP:WB, SAMPLE:FADU CELL,HEP2 CELL AND LSCC TISSUE, DILUTION:1:3000
Li Z,Feng J,Yang S,Meng P,Li J,Li H,Gao X,Zhang Y.Lipopolysaccharide-induced inflammation in human peritoneal mesothelial cells is controlled by ERK1/2-CDK5-PPARγ axis.Ann Transl Med.2021 May;9(10):850.doi:10.21037/atm-21-1623.PMID:34164484;PMCID:PMC8184493.
Species: Human
M00227-1 usage in article: APP:WB, SAMPLE:HMRSV5 CELL, DILUTION:1:10000
Jia Han,Wei Hou,Bi-qing Cai,Fan Zhang,Jian-cai Tang,"12-Epi-Napelline Inhibits Leukemia Cell Proliferation via the PI3K/AKT Signaling Pathway In Vitro and In Vivo",Evidence-Based Complementary and Alternative Medicine,vol.2021,Article ID 6687519,12 pages,2021.https://doi.org/10.1155/2021/6687519
Species: Human,Mouse
M00227-1 usage in article: APP:WB, SAMPLE:K-562 CELL, DILUTION:NA
Enoch Appiah Adu-Gyamfi,Jones Lamptey,Xue-Mei Chen,Fang-Fang Li,Cong Li,Ling-Ling Ruan,Xue-Niu Yang,Tai-Hang Liu,Ying-Xiong Wang,Yu-Bin Ding,Iodothyronine deiodinase 2 (DiO2) regulates trophoblast cell line cycle, invasion and apoptosis; and its downregulation is associated with early recurrent miscarriage,Placenta,Volume 111,2021,Pages 54-68,ISSN 0143-4004,https://doi.org/10.1016/j.placenta.2021.06.004.
Species: Human
M00227-1 usage in article: APP:WB, SAMPLE:HTR-8/SVNEO CELL, DILUTION:NA
Tibatan MA,Önay Uçar E.Long non-coding RNA NKILA regulates expression of HSP90α, NF-κB and β-catenin proteins in the MCF-7 breast cancer cell line.Mol Biol Rep.2021 Jun 14.doi:10.1007/s11033-021-06482-y.Epub ahead of print.PMID:34125329.
Species: Human
M00227-1 usage in article: APP:WB, SAMPLE:MCF-7 CELL, DILUTION:NA
Zhao,Y.,Jia,W.,Ren,S.,Xiao,W.,Li,G.,Jin,L.,& Lin,Y.(2021). Difluoromethylornithine attenuates isoproterenol‑induced cardiac hypertrophy by regulating apoptosis, autophagy and the mitochondria‑associated membranes pathway.Experimental and Therapeutic Medicine,22,870.https://doi.org/10.3892/etm.2021.10302
Species: Rat
M00227-1 usage in article: APP:WB, SAMPLE:VENTRICLE TISSUE, DILUTION:1:1000
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Customer Q&As
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17 Customer Q&As for Anti-GAPDH Rabbit Monoclonal Antibody
Question
Will anti-GAPDH Rabbit Monoclonal antibody M00227-1 work for Flow Cytometry with lymphoblast?
Verified Customer
Verified customer
Asked: 2020-03-30
Answer
According to the expression profile of lymphoblast, GAPDH is highly expressed in lymphoblast. So, it is likely that anti-GAPDH Rabbit Monoclonal antibody M00227-1 will work for Flow Cytometry with lymphoblast.
Boster Scientific Support
Answered: 2020-03-30
Question
I have a question about product M00227-1, anti-GAPDH Rabbit Monoclonal antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2020-02-24
Answer
We suggest not storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free M00227-1 anti-GAPDH Rabbit Monoclonal antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2020-02-24
Question
Is there a BSA free version of anti-GAPDH Rabbit Monoclonal antibody M00227-1 available?
Verified Customer
Verified customer
Asked: 2020-01-14
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-GAPDH Rabbit Monoclonal antibody M00227-1 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2020-01-14
Question
Is a blocking peptide available for product anti-GAPDH Rabbit Monoclonal antibody (M00227-1)?
Verified Customer
Verified customer
Asked: 2019-09-04
Answer
We do provide the blocking peptide for product anti-GAPDH Rabbit Monoclonal antibody (M00227-1). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-09-04
Question
Does anti-GAPDH Rabbit Monoclonal antibody M00227-1 work on canine Flow Cytometry with cervix carcinoma?
K. Mitchell
Verified customer
Asked: 2019-07-19
Answer
Our lab technicians have not validated anti-GAPDH Rabbit Monoclonal antibody M00227-1 on canine. You can run a BLAST between canine and the immunogen sequence of anti-GAPDH Rabbit Monoclonal antibody M00227-1 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated canine samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in canine cervix carcinoma in Flow Cytometry, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-07-19
Question
We have been able to see staining in rat muscle. Do you have any suggestions? Is anti-GAPDH Rabbit Monoclonal antibody supposed to stain muscle positively?
Verified Customer
Verified customer
Asked: 2019-07-03
Answer
Based on literature muscle does express GAPDH. Based on Uniprot.org, GAPDH is expressed in smooth muscle tissue, liver, lung, placenta, astrocytoma, leukemia, colon adenocarcinoma, eye, kidney, lung, lymph placenta, muscle, platelet, prostatic carcinoma, brain, cajal-retzius cell fetal brain cortex, heart, lymphoblast, cervix carcinoma, leukemic t-cell, cervix carcinoma erythroleukemia, among other tissues. Regarding which tissues have GAPDH expression, here are a few articles citing expression in various tissues:
Astrocytoma, Pubmed ID: 10944468
Cervix carcinoma, Pubmed ID: 17081983, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Heart, Pubmed ID: 7498159
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 2987855, 24275569
Lung, Pubmed ID: 3664468
Lymphoblast, Pubmed ID: 14654843
Muscle, Pubmed ID: 1193541, 11724794
Placenta, Pubmed ID: 1924305
Boster Scientific Support
Answered: 2019-07-03
Question
My boss were happy with the WB result of your anti-GAPDH Rabbit Monoclonal antibody. However we have observed positive staining in cervix carcinoma erythroleukemia cytosol using this antibody. Is that expected? Could you tell me where is GAPDH supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-06-18
Answer
From literature, cervix carcinoma erythroleukemia does express GAPDH. Generally GAPDH expresses in cytoplasm, cytosol. Regarding which tissues have GAPDH expression, here are a few articles citing expression in various tissues:
Astrocytoma, Pubmed ID: 10944468
Cervix carcinoma, Pubmed ID: 17081983, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Heart, Pubmed ID: 7498159
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 2987855, 24275569
Lung, Pubmed ID: 3664468
Lymphoblast, Pubmed ID: 14654843
Muscle, Pubmed ID: 1193541, 11724794
Placenta, Pubmed ID: 1924305
Boster Scientific Support
Answered: 2019-06-18
Question
I was wanting to use your anti-GAPDH Rabbit Monoclonal antibody for Flow Cytometry for cow lymphoblast on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for cow lymphoblast identification?
Verified Customer
Verified customer
Asked: 2019-02-11
Answer
You can see on the product datasheet, M00227-1 anti-GAPDH Rabbit Monoclonal antibody has been tested for Flow Cytometry, IF, IHC, ICC, WB on canine, chicken, cow, human, monkey, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in cow lymphoblast in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-02-11
Question
I see that the anti-GAPDH Rabbit Monoclonal antibody M00227-1 works with Flow Cytometry, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-01-16
Answer
You can find protocols for Flow Cytometry on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2019-01-16
Question
Thanks for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for lymphoblast using anti-GAPDH Rabbit Monoclonal antibody M00227-1. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2018-05-30
Answer
Thanks for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-05-30
Question
We are currently using anti-GAPDH Rabbit Monoclonal antibody M00227-1 for chicken tissue, and we are content with the Flow Cytometry results. The species of reactivity given in the datasheet says canine, chicken, cow, human, monkey, mouse, rat. Is it possible that the antibody can work on monkey tissues as well?
B. Jones
Verified customer
Asked: 2018-04-04
Answer
The anti-GAPDH Rabbit Monoclonal antibody (M00227-1) has not been validated for cross reactivity specifically with monkey tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in monkey you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2018-04-04
Question
We want to test anti-GAPDH Rabbit Monoclonal antibody M00227-1 on cow lymphoblast for research purposes, then I may be interested in using anti-GAPDH Rabbit Monoclonal antibody M00227-1 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2018-04-02
Answer
The products we sell, including anti-GAPDH Rabbit Monoclonal antibody M00227-1, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2018-04-02
Question
We are interested in using your anti-GAPDH Rabbit Monoclonal antibody for antimicrobial humoral immune response mediated by antimicrobial peptide studies. Has this antibody been tested with western blotting on hela cell lysate? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2018-03-09
Answer
We appreciate your inquiry. This M00227-1 anti-GAPDH Rabbit Monoclonal antibody is validated on hela cell lysate. It is guaranteed to work for Flow Cytometry, IF, IHC, ICC, WB in canine, chicken, cow, human, monkey, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2018-03-09
Question
See attached the WB image, lot number and protocol we used for lymphoblast using anti-GAPDH Rabbit Monoclonal antibody M00227-1. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2018-01-05
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-01-05
Question
Will M00227-1 anti-GAPDH Rabbit Monoclonal antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2017-09-27
Answer
As indicated on the product datasheet, M00227-1 anti-GAPDH Rabbit Monoclonal antibody as been tested on Flow Cytometry. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2017-09-27
Question
We bought anti-GAPDH Rabbit Monoclonal antibody for IHC on leukemia in the past. I am using canine, and We are going to use the antibody for Flow Cytometry next. I am interested in examining leukemia as well as lymphoblast in our next experiment. Do you have any suggestion on which antibody would work the best for Flow Cytometry?
M. Wu
Verified customer
Asked: 2015-06-17
Answer
I have checked the website and datasheets of our anti-GAPDH Rabbit Monoclonal antibody and I see that M00227-1 has been tested on canine in both IHC and Flow Cytometry. Thus M00227-1 should work for your application. Our Boster satisfaction guarantee will cover this product for Flow Cytometry in canine even if the specific tissue type has not been validated. We do have a comprehensive range of products for Flow Cytometry detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2015-06-17
Question
Is this M00227-1 anti-GAPDH Rabbit Monoclonal antibody reactive to the isotypes of GAPDH?
H. Li
Verified customer
Asked: 2014-09-15
Answer
The immunogen of M00227-1 anti-GAPDH Rabbit Monoclonal antibody is A synthesized peptide derived from human GAPDH. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2014-09-15