Human/Mouse/Rat GAPDH Antibody

Catalog # Availability Size / Price Qty
AF5718
AF5718-SP

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Detection of Human/Mouse/Rat GAPDH/G3PDH by Western Blot.
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Product Details
Citations (25)
FAQs
Supplemental Products
Reviews (5)

Human/Mouse/Rat GAPDH Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat GAPDH/G3PDH in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human GAPDH-2 is observed. 
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human GAPDH/G3PDH
Met1-Ala150
Accession # P04406
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Immunocytochemistry
5-25 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human/Mouse/Rat GAPDH/G3PDH antibody by Western Blot. View Larger

Detection of Human/Mouse/Rat GAPDH/G3PDH by Western Blot. Western blot shows lysates of mouse, human, and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat GAPDH/G3PDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GAPDH/G3PDH at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Immunocytochemistry GAPDH antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

GAPDH in HeLa Human Cell Line. GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry GAPDH/G3PDH antibody in Human Kidney by Immunohistochemistry (IHC-P). View Larger

GAPDH/G3PDH in Human Kidney. GAPDH/G3PDH was detected in immersion fixed paraffin-embedded sections of normal human kidney using Goat Anti-Human/Mouse/Rat GAPDH/G3PDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 8 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei of convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.This application has not been tested in mouse or rat samples.

Immunohistochemistry GAPDH antibody in Human Prostate Cancer Tissue by Immunohistochemistry (IHC-P). View Larger

GAPDH in Human Prostate Cancer Tissue. GAPDH was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Goat Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western Detection of Human GAPDH antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Human GAPDH by Simple WesternTM. Simple Western lane view shows lysates of human brain (motor cortex) tissue and HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for GAPDH at approximately 43 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Detection of Mouse GAPDH by Western Blot View Larger

Detection of Mouse GAPDH by Western Blot Role of Akt pathway in aliskiren's effects on attenuating collagen expression.A and B demonstrate the results of Western blotting for phospho- and total Akt in hearts of mice treated with various diets. C shows changes in the protein levels of collagen type I, phospho- and total Akt in response to 24 hours of exposure to D,L. Homocysteine (200 µmol/L) with or without pre-treatment with aliskiren 10 µmol/L (AL), wortmannin 1 µmol/L or Akt Inhibitor VIII 1 µmol/L. Hhe – Hhe-inducing diet; D,L, homocysteine (lower panel); AL 0.05– aliskiren 0.5 mg/kg/day; AL 5– aliskiren 5 mg/kg/day; AL 50– aliskiren 50 mg/kg/day; Wort – wortmannin; Akt in – Akt inhibitor VIII. *p<0.05; **p<0.01; and ***p<0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24349097), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Rat GAPDH by Western Blot View Larger

Detection of Rat GAPDH by Western Blot Effect of prodigiosins on the mRNA and protein expression of HO-1 in gastric tissue.Data (means ± SEM of three assays) were normalized to the levels of GAPDH and expressed as fold induction (log2 scale), relative to the level of HO-1 in the control. aP < 0.05, significant difference vs. the health rats; bP < 0.05, significant difference vs. HCl/ethanol-induced rats, Duncan's post-hoc test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31194753), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Human GAPDH by Western Blot View Larger

Detection of Human GAPDH by Western Blot The DU145FP cells express higher levels of cancer stemness related genes. (A) Transcriptome analyses of prostate cancer stemness-related markers. The Log2 transformed ratio of FPKM values (e.g., FP/WT) are indicated by color-coded index bars. (B) Cells were stained with fluorophore conjugated monoclonal antibodies against CD44 and EPCAM, the antibody against integrin alpha 2 beta 3 without fluorophore conjugation was detected by using secondary antibody conjugated with fluorophore, and stained cells were analyzed by flow cytometry. The scatter plots and histograms show representative results. CTRL denote cell stained with isotype control antibody or only secondary antibody; ST denote cell stained with antibody against EPCAM and integrin alpha 2 beta 3; US denote unstained cells. (C) Immunoblotting confirmation of the protein expression of A-tubulin, GAPDH, SOX9, SLUG, HIF1A, HIF2A, CD44 and integrin alpha 2 beta 3. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28698547), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Rat GAPDH by Western Blot View Larger

Detection of Rat GAPDH by Western Blot Effects of aliskiren on homocysteine-induced matrix expression in cultured cardiac fibroblasts.Fibroblasts were cultured in the presence or absence of D,L, homocysteine 200 µmol/L for 24 hours after pretreatment with varying concentrations of aliskiren (0.1–10 µmol/L). Real time PCR was done to measure relative expression of COLIA2 (alpha 2 chain of type I collagen; A) and COL3A1 (alpha 1 chain of type III collagen; B) genes. Figure 5C demonstrates the results of real time PCR to compare the effects of losartan and captopril with aliskiren on homocysteine induced expression of COL1A2 gene. Western blotting was conducted to measure the expression of collagen type I protein (D and E) Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) was used as internal control. Hcy-D,L, homocysteine; AL-aliskiren; Los – losartan; and Cap – captopril. *p<0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24349097), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Mouse GAPDH by Western Blot View Larger

Detection of Mouse GAPDH by Western Blot Role of Akt pathway in aliskiren's effects on attenuating collagen expression.A and B demonstrate the results of Western blotting for phospho- and total Akt in hearts of mice treated with various diets. C shows changes in the protein levels of collagen type I, phospho- and total Akt in response to 24 hours of exposure to D,L. Homocysteine (200 µmol/L) with or without pre-treatment with aliskiren 10 µmol/L (AL), wortmannin 1 µmol/L or Akt Inhibitor VIII 1 µmol/L. Hhe – Hhe-inducing diet; D,L, homocysteine (lower panel); AL 0.05– aliskiren 0.5 mg/kg/day; AL 5– aliskiren 5 mg/kg/day; AL 50– aliskiren 50 mg/kg/day; Wort – wortmannin; Akt in – Akt inhibitor VIII. *p<0.05; **p<0.01; and ***p<0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24349097), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Mouse GAPDH by Western Blot View Larger

Detection of Mouse GAPDH by Western Blot Effects of aliskiren on myocardial matrix gene expression.Real time PCR was performed with ventricular tissue obtained from mice after 12-inducing diet and different doses of aliskiren. Figure 3A demonstrates changes in expression of COL1A1(alpha 1 chain of type I collagen) and 3B shows changes in COL1A2(alpha2 chain of type I collagen) genes. Lower right panels (3C and 3D) demonstrate the results of western blotting for collagen type I in various groups. Hhe – Hhe-inducing diet; AL 0.5– aliskiren 0.5 mg/kg/day; AL 5– aliskiren 5 mg/kg/day; and AL 50– aliskiren 50 mg/kg/day. * p<0.05; and *** p<0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24349097), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: GAPDH

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a 36‑40 kDa member of the GAPDH family of enzymes. It is a widely expressed heterotetramer that is found in both the nucleus and cytoplasm. Although GAPDH was initially identified as a glycolytic enzyme that converted G3P into 1,3 diphosphoglycerate, it is now recognized to participate in no less than endocytosis, membrane fusion, vesicular secretory transport, DNA replication and repair, and apoptosis. Human GAPDH is 335 amino acids (aa) in length and contains two NAD binding sites (Asp35 and Asn316) with a catalytic region between aa 151‑155. GAPDH contains more than 19 posttranslational modifications, including methylation, deamidation and phosphorylation. One splice variant shows a 10 aa substitution for aa 319‑335. Over amino acid 1‑150, human GAPDH shares 92% aa identity with mouse GAPDH.

Long Name
Glyceraldehyde-3-phosphate Dehydrogenase
Entrez Gene IDs
2597 (Human); 14433 (Mouse); 24383 (Rat)
Alternate Names
aging-associated gene 9 protein; EC 1.2.1; EC 1.2.1.12; EC 2.6.99.-; G3PD; G3PDH; GAPD; GAPDH; glyceraldehyde 3-phosphate dehydrogenase; glyceraldehyde-3-phosphate dehydrogenase; MGC88685; Peptidyl-cysteine S-nitrosylase GAPDH

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Citations for Human/Mouse/Rat GAPDH Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

25 Citations: Showing 1 - 10
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  1. Discovery and Characterization of ZL-2201, a Potent, Highly Selective, and Orally Bioavailable Small-molecule DNA-PK Inhibitor.
    Authors: Lal S, Bhola N H et al.
    Cancer Res Commun
    Species: Human
    Sample Types:
    Applications: Simple Western
  2. Metformin Mitigated Obesity-Driven Cancer Aggressiveness in Tumor-Bearing Mice
    Authors: CJ Chen, CC Wu, CY Chang, JR Li, YC Ou, WY Chen, SL Liao, JD Wang
    International Journal of Molecular Sciences, 2022-08-15;23(16):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: Western Blot
  3. Robo4 is constitutively shed by ADAMs from endothelial cells and the shed Robo4 functions to inhibit Slit3-induced angiogenesis
    Authors: W Xiao, A Pinilla-Ba, J Faulkner, X Song, P Prabhakar, H Qiu, KW Moremen, A Ludwig, PJ Dempsey, P Azadi, L Wang
    Scientific Reports, 2022-03-14;12(1):4352.
    Species: Human, Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. Decidua Parietalis Mesenchymal Stem/Stromal Cells and Their Secretome Diminish the Oncogenic Properties of MDA231 Cells In Vitro
    Authors: Y Basmaeil, E Bahattab, A Al Subayyi, HB Kulayb, M Alrodayyan, M Abumaree, T Khatlani
    Cells, 2021-12-10;10(12):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. Therapeutic Hypothermia Inhibits the Classical Complement Pathway in a Rat Model of Neonatal Hypoxic-Ischemic Encephalopathy
    Authors: TA Shah, HK Pallera, CL Kaszowski, WT Bass, FA Lattanzio
    Frontiers in Neuroscience, 2021-02-12;15(0):616734.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. Multiplex coherent anti-Stokes Raman scattering microspectroscopy detection of lipid droplets in cancer cells expressing TrkB
    Authors: T Guerenne-D, V Couderc, L Duponchel, V Sol, P Leproux, JM Petit
    Sci Rep, 2020-10-07;10(1):16749.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  7. Overexpression of microRNA-367 inhibits angiogenesis in ovarian cancer by downregulating the expression of LPA1
    Authors: Q Zheng, X Dai, W Fang, Y Zheng, J Zhang, Y Liu, D Gu
    Cancer Cell Int, 2020-10-02;20(0):476.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  8. Mouse WIF1 Is Only Modified with O-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites
    Authors: F Pennarubia, E Pinault, B Al Jaam, CE Brun, A Maftah, A Germot, S Legardinie
    Biomolecules, 2020-08-28;10(9):.
    Species: Hamster
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  9. scAAV2-Mediated C3 Transferase Gene Therapy in a Rat Model with Retinal Ischemia/Reperfusion Injuries
    Authors: J Tan, X Zhang, D Li, G Liu, Y Wang, D Zhang, X Wang, W Tian, X Dong, L Zhou, X Zhu, X Liu, N Fan
    Mol Ther Methods Clin Dev, 2020-04-25;17(0):894-903.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  10. C3 Transferase-Expressing scAAV2 Transduces Ocular Anterior Segment Tissues and Lowers Intraocular Pressure in Mouse and Monkey
    Authors: J Tan, X Wang, S Cai, F He, D Zhang, D Li, X Zhu, L Zhou, N Fan, X Liu
    Mol Ther Methods Clin Dev, 2019-11-29;17(0):143-155.
    Species: Human
    Sample Types: Cell
    Applications: Western Blot
  11. PDHA1 Gene Knockout In Human Esophageal Squamous Cancer Cells Resulted In Greater Warburg Effect And Aggressive Features In Vitro And In Vivo
    Authors: L Liu, J Cao, J Zhao, X Li, Z Suo, H Li
    Onco Targets Ther, 2019-11-18;12(0):9899-9913.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  12. Downregulation of POFUT1 Impairs Secondary Myogenic Fusion Through a Reduced NFATc2/IL-4 Signaling Pathway
    Authors: A Der Vartan, J Chabanais, C Carrion, A Maftah, A Germot
    Int J Mol Sci, 2019-09-06;20(18):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  13. Prodigiosins from a marine sponge-associated actinomycete attenuate HCl/ethanol-induced gastric lesion via antioxidant and anti-inflammatory mechanisms
    Authors: MS Abdelfatta, MIY Elmallah, HY Ebrahim, RS Almeer, RMA Eltanany, AE Abdel Mone
    PLoS ONE, 2019-06-13;14(6):e0216737.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  14. Cysteinyl leukotriene receptor 2 drives lung immunopathology through a platelet and high mobility box 1-dependent mechanism
    Authors: T Liu, NA Barrett, Y Kanaoka, K Buchheit, TM Laidlaw, D Garofalo, J Lai, HR Katz, C Feng, JA Boyce
    Mucosal Immunol, 2019-01-21;0(0):.
    Species: Mouse
    Sample Types: Tissue Lysates
    Applications: Western Blot
  15. Regulatory Evolution Drives Evasion of Host Inflammasomes by Salmonella Typhimurium
    Authors: B Ilyas, DT Mulder, DJ Little, W Elhenawy, MM Banda, D Pérez-Mora, CN Tsai, NYE Chau, VH Bustamante, BK Coombes
    Cell Rep, 2018-10-23;25(4):825-832.e5.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  16. Alterations in Adiposity and Glucose Homeostasis in Adult Gasp-1 Overexpressing Mice
    Authors: L Périè, A Parenté, F Baraige, L Magnol, V Blanquet
    Cell. Physiol. Biochem., 2017-12-08;44(5):1896-1911.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  17. Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Authors: MK Shim, HY Yoon, S Lee, MK Jo, J Park, JH Kim, SY Jeong, IC Kwon, K Kim
    Sci Rep, 2017-11-30;7(1):16635.
    Species: Human
    Sample Types: Whole Cells
    Applications: Western Blot
  18. Metabolic reprogramming is associated with flavopiridol resistance in prostate cancer DU145 cells
    Authors: X Li, J Lu, Q Kan, X Li, Q Fan, Y Li, R Huang, A Slipicevic, HP Dong, L Eide, J Wang, H Zhang, V Berge, MA Goscinski, G Kvalheim, JM Nesland, Z Suo
    Sci Rep, 2017-07-11;7(1):5081.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  19. Salmonella Typhimurium disrupts Sirt1/AMPK checkpoint control of mTOR to impair autophagy
    Authors: R Ganesan, NJ Hos, S Gutierrez, J Fischer, JM Stepek, E Daglidu, M Krönke, N Robinson
    PLoS Pathog, 2017-02-13;13(2):e1006227.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  20. Activation of KLF1 Enhances the Differentiation and Maturation of Red Blood Cells from Human Pluripotent Stem Cells
    Authors: CT Yang, R Ma, RA Axton, M Jackson, AH Taylor, A Fidanza, L Marenah, J Frayne, JC Mountford, LM Forrester
    Stem Cells, 2017-01-19;35(4):886-897.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  21. Up-regulation of VEGF by retinoic acid during hyperoxia prevents retinal neovascularization and retinopathy.
    Authors: Wang L, Shi P, Xu Z, Li J, Xie Y, Mitton K, Drenser K, Yan Q
    Invest Ophthalmol Vis Sci, 2014-05-27;55(7):4276-87.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  22. Prenatal retinoid deficiency leads to airway hyperresponsiveness in adult mice.
    Authors: Chen, Felicia, Marquez, Hector, Kim, Youn-Kyu, Qian, Jun, Shao, Fengzhi, Fine, Alan, Cruikshank, William, Quadro, Loredana, Cardoso, Wellingt
    J Clin Invest, 2014-01-09;124(2):801-11.
    Species: Mouse
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  23. Effects of direct Renin inhibition on myocardial fibrosis and cardiac fibroblast function.
    Authors: Zhi, Hui, Luptak, Ivan, Alreja, Gaurav, Shi, Jianru, Guan, Jian, Metes-Kosik, Nicole, Joseph, Jacob
    PLoS ONE, 2013-12-11;8(12):e81612.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot
  24. The hypoxic microenvironment upgrades stem-like properties of ovarian cancer cells.
    Authors: Liang D, Ma Y, Liu J, Trope CG, Holm R, Nesland JM, Suo Z
    BMC Cancer, 2012-05-29;12(0):201.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  25. Overexpression of carbonic anhydrase and HIF-1alpha in Wilms tumours.
    Authors: Dungwa JV, Hunt LP, Ramani P
    BMC Cancer, 2011-09-12;11(0):390.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: Western Blot

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Reviews for Human/Mouse/Rat GAPDH Antibody

Average Rating: 4.8 (Based on 5 Reviews)

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Human/Mouse/Rat GAPDH Antibody
By Anonymous on 09/28/2020
Application: Simple Western Sample Tested: HT-29 human colon adenocarcinoma cell line Species: Human

Human/Mouse/Rat GAPDH Antibody
By Anonymous on 10/17/2018
Application: WB Sample Tested: 4T1 mouse breast cancer cell line,EMT6 cells Species: Mouse

Human/Mouse/Rat GAPDH Antibody
By Anonymous on 07/21/2018
Application: WB Sample Tested: HEK293 human embryonic kidney cell line Species: Human

Human/Mouse/Rat GAPDH Antibody
By Anonymous on 01/25/2018
Application: WB Sample Tested: IPS2 induced pluripotent stem cells Species: Mouse

Human/Mouse/Rat GAPDH Antibody
By Kimberly Schwartz on 05/16/2017
Application: WB Sample Tested: Lysate and purified protein Species: Human

The blot contains HeLa cell lysate (1, 2, 3 and 5 µg) spiked with 200 ng of human transferrin per lane. Transferrin is detected in the red channel, tubulin is detected in the green channel and GAPDH is detected in the blue channel.