GAPDH Monoclonal Antibody

Code CSB-MA000071M0m
Size US$200Purchase it in Cusabio online store
(only available for customers from the US)
Image
  • Western Blot
    Positive WB detected in: 15μg hela whole cell lysate
    GAPDH antibody at 1:100000, 1:200000, 1:400000, 1:800000, 1:1600000
    Secondary
    Goat polyclonal to mouse IgG at 1/50000 dilution
    Predicted band size: 36 KDa
    Observed band size: 36 KDa
    Exposure time: 5min

  • Western Blot
    Positive WB detected in: Hela whole cell lysate at 10μg, 5μg, 2.5μg, 1.25μg, 0.625μg, 0.3125μg
    All lanes: GAPDH antibody at 1:5000
    Secondary
    Goat polyclonal to mouse IgG at 1/50000 dilution
    Predicted band size: 36 KDa
    Observed band size: 36 KDa
    Exposure time: 5min

  • Western Blot
    Positive WB detected in: Hela whole cell lysate, HepG2 whole cell lysate, Jurkat whole cell lysate, MCF-7 whole cell lysate
    All lanes: GAPDH antibody at 1:2000
    Secondary
    Goat polyclonal to mouse IgG at 1/50000 dilution
    Predicted band size: 36 KDa
    Observed band size: 36 KDa
    Exposure time: 30s

  • Western Blot
    Positive WB detected in: U87 whole cell lysate, PC3 whole cell lysate, 293 whole cell lysate, U251 whole cell lysate, A549 whole cell lysate, A375 whole cell lysate, MG-63 whole cell lysate
    All lanes GAPDH antibody at 1:5000
    Secondary
    Goat polyclonal to mouse IgG at 1/50000 dilution
    Predicted band size: 36 KDa
    Observed band size: 36 KDa
    Exposure time: 30s

  • Western Blot
    Positive WB detected in: Rat heart tissue, Rat kidney tissue, Rat skeletal muscle tissue, Rat liver tissue, Rat brain tissue tissue, Rat spleen tissue
    All lanes GAPDH antibody at 1:1000
    Secondary
    Goat polyclonal to mouse IgG at 1/50000 dilution
    Predicted band size: 36 KDa
    Observed band size: 36 KDa
    Exposure time: 1min

  • Western Blot
    Positive WB detected in: Rabbit heart tissue, Rabbit liver tissue,Rabbit spleen tissue, Rabbit lung tissue, Rabbit kidney tissue, Rabbit small intestine tissue, Rabbit skeletal muscle tissue
    All lanes GAPDH antibody at 1:5000
    Secondary
    Goat polyclonal to mouse IgG at 1/50000 dilution
    Predicted band size: 36 KDa
    Observed band size: 36 KDa
    Exposure time: 5min

  • IHC image of CSB-MA000071M0m diluted at 1:100 and staining in paraffin-embedded human colon cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

  • IHC image of CSB-MA000071M0m diluted at 1:100 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

  • IHC image of CSB-MA000071M0m diluted at 1:100 and staining in paraffin-embedded human kidney tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

  • Immunofluorescence staining of Hela cells with CSB-MA000071M0m at 1:220, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

  • Immunofluorescence staining of HepG2 cells with CSB-MA000071M0m at 1:220, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

  • Immunoprecipitating GAPDH in Hela whole cell lysate
    Lane 1: Mouse control IgG instead of CSB-MA000071M0m in Hela whole cell lysate.
    Lane 2: CSB-MA000071M0m (1μl) + Hela whole cell lysate (500μg)
    Lane 3: Hela whole cell lysate (20μg)
    For western blotting, the blot was detected with CSB-MA000071M0m at 1:5000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:2000

  • Overlay histogram showing Hela cells stained with CSB-MA000071M0m (red line). The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1:200/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/100 dilution for 30min at 4°C. Isotype control antibody (green line) was mouse IgG1 (1:200/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.

  • Overlay histogram showing Jurkat cells stained with CSB-MA000071M0m (red line). The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1:200/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/100 dilution for 30min at 4°C. Isotype control antibody (green line) was mouse IgG1 (1:200/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.

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Product Details

Full Product Name Mouse anti-Human GAPDH Monoclonal antibody
Uniprot No. P04406
Target Names GAPDH
Alternative Names GAPDH; G3PD; GAPD; MGC88685
Raised in Mouse
Species Reactivity Human, Rat, Rabbit
Immunogen Recombinant Human GAPDH protein (3-335AA)
Immunogen Species Human
Conjugate Non-conjugated
Clonality Monoclonal
Isotype IgG1
Clone No. 14C2F11
Purification Method >95%, Protein G purified
Concentration It differs from different batches. Please contact us to confirm it.
Buffer Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Form Liquid
Tested Applications ELISA, WB, IHC, IP, IF, FC
Recommended Dilution
Application Recommended Dilution
WB 1:5000-1:1600000
IHC 1:50-1:500
IF 1:50-1:200
IP 1µl-2µl
FC 1:100-1:300
Protocols ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunoprecipitation (IP) Protocol
Immunofluorescence (IF) Protocol
Flow Cytometry (FC) Protocol
Troubleshooting and FAQs Antibody FAQs
Storage Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
Lead Time Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Citations

Concurrent EPA and DHA Supplementation Impairs Brown Adipogenesis of C2C12 Cells. S Ghnaimawi,Frontiers in Genetics,2020

Applications: Western Blot
Antibody dilution factor: 1:1000
Review: Protein level measurement by western blot of myogenesis regulating genes in C2C12 and GAPDH as housekeeping gene n = 4.
PMID: 32595696

Quadrella incana (Capparaceae) Leaf Extract Enhances Proliferation and Maintenance of Neural Stem/Progenitor Cells through Upregulating Glycolytic Flux and Redox Potential. M Kang,Oxid Med Cell Longev,2020

Applications: Western blot
Antibody dilution factor: 1:1,000
Review: Western blot analysis of Bmi1, βIII tubulin, and phospho-Histone 3 (Ser 10) in NSPCs kept under proliferating media upon treatment for 3 days with DMSO, Quadrella incana leaf extract, or six different plant crude extracts. GAPDH was used as a loading control. GAPDH was used as a loading control.
PMID: 32411330

Emerin suppresses Notch signaling by restricting the Notch intracellular domain to the nuclear membrane. B Lee.et al,Biochimica et biophysica acta-molecular cell research,2017

Applications: WB
Antibody dilution factor: 1: 5000
Review: HeLa cells were treated with si BAF #2 or si Control for 72 h. Cells were fractionated and subjected to western blot analyses (upper panel). Fractionation was verified by using Lamin A/C antibody (nuclear fraction) or GAPDH antibody (cytosolic fraction). The intensity of the cytosolic emerin was analyzed using ImageJ software (lower panel).
PMID: 27865926

A fluorescence method for determination of glucose transport by intestinal BBMV of common carp. Yang LP.et al,Anal Biochem,2017

Application: Immunoblot analyses
Review: Immunoblot analyses show the presence of SGLT-1 on BBMV of common carp.
PMID: 28847591

Customer Reviews and Q&A

 Customer Reviews
Average Rating:
4.7 - 7 reviews

Submit a Review here

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Applications : Western Blot (WB)

Sample type: Human ovarian granulosa cell carcinoma

Sample dilution: 1:1200

Review: Experiment success, antibody is usable!

By Anonymous

Applications : Western Blot (WB)

Sample type: KELLY

Sample dilution: 1:1000

Review: GAPDH has good results in this experiment, with correct size and single band.

By Anonymous

Applications : Western Blot (WB)

Sample type: Human ovarian granulosa cell carcinoma

Sample dilution: 1:1000

Review: The experiment was successful, and the antibody was successfully detected on human cells, which meets the experimental requirements and can be purchased and used in the future.

By Anonymous

Applications : Western Blot (WB)

Sample type: Rat kidney tissue

Sample dilution: 1:5000

Review: Good antibody, good titer.

By Anonymous

Applications : Western Blot (WB)

Sample type: Mouse

Sample dilution: 1:5000

Review: Antibody specificity is very good.

By Anonymous

Applications : Western Blot (WB)

Sample type: Human 231 cell

Sample dilution: 1:2000

Review: The destination band is correct and single.

By Anonymous

Applications : Western Blot (WB)

Sample type: Cell Lysate (Chicken)

Sample dilution: 1:2000

Review: Incubate at 4 °C overnight with 1: 2000 dilution ratio, the bands are clear, the specificity is good, and it can be recycled and reused with good results.

By Anonymous

Target Data

Function Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.
Gene References into Functions
  1. This suggests that RX624 might be useful as a drug against polyglutamine pathologies, and that is could be administered exogenously without affecting target cell physiology. This protective effect was validated by the similar effect of an anti-GAPDH specific antibody. PMID: 28450110
  2. GAPDH can interact with proteins participating in DNA repair, such as APE1, PARP1, HMGB1, and HMGB2. In this review, the functions of GAPDH associated with DNA repair are discussed in detail. PMID: 28601074
  3. Nitric oxide-induced GAPDH aggregation specifically induces mitochondrial dysfunction via permeability transition pore opening, leading to cell death. PMID: 28167533
  4. GAPDH may act as a chaperone in heme transfer to downstream areas PMID: 28315300
  5. NAD(+) inhibited both GAPDH aggregation and co-aggregation with GOSPEL, a hitherto undescribed effect of the coenzyme against the consequences of oxidative stress. PMID: 27282776
  6. Monoclonal Antibodies DSHB-hGAPDH-2G7 and DSHB-hGAPDH-4B7 Against Human Glyceraldehyde-3-Phosphate Dehydrogenase. PMID: 27556912
  7. the present study suggests that GAPDH plays an important role in cancer metastasis by affecting EMT through regulation of Sp1-mediated SNAIL expression. PMID: 27878251
  8. Knockdown of LAMP2A, a CMA-related protein, and TSG101, an mA-related protein, significantly but only partially decreased the punctate accumulation of GAPDH-HT in AD293 cells and primary cultured rat cortical neurons. PMID: 27377049
  9. In conclusion, the data show that two GAPDH binders could be therapeutically relevant in the treatment of injuries stemming from hard oxidative stress. PMID: 26748070
  10. transient silencing of GAPDH reduces intracellular ROS and facilitates increased autophagy, thereby reducing acute hypoxia and reoxygenation injury as well as the resulting apoptosis and necrosis. PMID: 26279122
  11. This review will summarize our current understanding of GAPDH-mediated regulation of RNA function PMID: 26564736
  12. analysis of PSCA level in the peripheral blood of PC patients who underwent radical prostatectomy shows it is related to a GADPH reference level (PSCA/GAPDH ratio) PMID: 26527100
  13. In 60 % of patients with type 2 diabetes, a reversible inhibition of GAPDH is observed. PMID: 25189828
  14. The results of this study led us to conclude that in cancer cells constantly exposed to conditions of oxidative stress, the protective power of Hsp70 should be abolished by specific inhibitors of Hsp70 expression. PMID: 26713364
  15. GAPDH and protoporphyrinogen oxidase were shown to have higher expression in faster growing cell lines and primary tumors. Pharmacologic inhibition of GAPDH or PPOX reduced the growth of colon cancer cells in vitro PMID: 25944804
  16. The levels of GAPDH protein were significantly up-regulated in lung squamous cell carcinoma tissues and elevated GAPDH expression is associated with the proliferation and invasion of lung and esophageal squamous cell carcinomas. PMID: 25944651
  17. Genetic variants in GAPDH confer susceptibility to sporadic Parkinson's disease in a Chinese Han population. PMID: 26258539
  18. Data revealed that GAPDH is a phosphorylation substrate for AMPK and its interaction with Sirt1 in the nucleus. The phosphorylation and the nuclear translocation of GAPDH mediate rapid Sirt1 activation and autophagy initiation under glucose deprivation. PMID: 26626483
  19. findings demonstrate that dissociation of the GAPDH/Siah1 pro-apoptotic complex can block high glucose-induced pericyte apoptosis, widely considered a hallmark feature of diabetic retinopathy PMID: 26438826
  20. Extracellular GAPDH, or its N-terminal domain, inhibited gastric cancer cell growth. GAPDH bound to E-cadherin and downregulated the mTOR-p70S6 kinase pathway. PMID: 25785838
  21. suggests that GAPDH aggregates accelerate Abeta amyloidogenesis, subsequently leading to mitochondrial dysfunction and neuronal cell death in the pathogenesis of AD PMID: 26359500
  22. The level of GAPDH-AP DNA adduct formation depends on oxidation of the protein SH-groups; disulfide bond reduction in GAPDH leads to the loss of its ability to form the adducts with AP DNA PMID: 26203648
  23. The activity of GAPDS was significantly positively correlated with sperm motility and negatively with the incidence of infertility. PMID: 26255202
  24. The N terminus of nuclear GAPDH binds with PARP-1, and this complex promotes PARP-1 overactivation both in vitro and in vivo. PMID: 25882840
  25. deregulated GAPDH expression promotes NF-kappaB-dependent induction of HIF-1alpha and has a key role in lymphoma vascularization and aggressiveness PMID: 25394713
  26. analysis of how flux through GAPDH is a limiting step in aerobic glycolysis PMID: 25009227
  27. astrocytic production of D-serine is modulated by glycolytic activity via interactions between GAPDH and SRR. PMID: 25870284
  28. Dimer and tetramer interface residues in adenine-uridine rich elements are important for GAPDH-RNA binding. PMID: 25451934
  29. Siah1 is a substrate of ASK1 for activation of the GAPDH-Siah1 oxidative stress signaling cascade. PMID: 25391652
  30. GAPDH expression is deregulated during melanoma progression. PMID: 25550585
  31. Oxidation of an exposed methionine instigates the aggregation of glyceraldehyde-3-phosphate dehydrogenase. PMID: 25086035
  32. MZF-1 binds to and positively regulates the GAPDH promoter, indicating a role for GAPDH in calcitriol-mediated signaling. PMID: 25065746
  33. The protein encoded by this gene contains a peptide that displays antimicrobial activity against E. coli, P. aeruginosa, and C. albicans. PMID: 22832495
  34. GAPDH gene over expression in resected tumor samples is an adverse prognostic factor in non small cell lung cancer. PMID: 23988223
  35. This review describes the structure and localization of GAPDH in cells as well as the latest discoveries on the multifunctional properties of the enzyme. PMID: 24018444
  36. TG2-dependent GAPDH deamidation was suggested to participate in actin cytoskeletal remodeling. PMID: 24375405
  37. acetylation of GAPDH (K254) is reversibly regulated by the acetyltransferase PCAF and the deacetylase HDAC5. PMID: 24362262
  38. GAPDH binds to active Akt, leading to Bcl-xL increase and escape from caspase-independent cell death. PMID: 23645209
  39. GAPDH is a moonlighting protein that functions as a glycolytic enzyme as well as a uracil DNA glycosylase. PMID: 20727968
  40. Results indicate that CIB1 is uniquely positioned to regulate PI3K/AKT and MEK/ERK signaling and that simultaneous disruption of these pathways synergistically induces a nuclear GAPDH-dependent cell death. PMID: 22964641
  41. The data presented demonstrate that up-regulation of GAPDH positively associated genes is proportional to the malignant stage of various tumors and is associated with an unfavourable prognosis. PMID: 23620736
  42. In a yeast two-hybrid screen of a heart cDNA library with Mst1 as bait, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was identified as an Mst1-interacting protein. PMID: 23527007
  43. interaction between prolyl oligopeptidase and glyceraldehyde-3-phosphate dehydrogenase is required for cytosine arabinoside-induced glyceraldehyde-3-phosphate dehydrogenase nuclear translocation and cell death PMID: 23348613
  44. NleB, a bacterial glycosyltransferase, targets GAPDH function to inhibit NF-kappaB activation. PMID: 23332158
  45. GAPDH binds to alkylated, single-stranded, double-stranded and telomeric sequences in a drug-dependent and DNA sequence/structure-dependent manner. PMID: 23409959
  46. GAPDH negatively regulates HIV-1 infection and provide insights into a novel function of GAPDH in the HIV-1 life cycle and a new host defense mechanism against HIV-1 infection. PMID: 23237566
  47. The strength, selectivity, reversibility, and redox sensitivity of heme binding to GAPDH are consistent with it performing heme sensing or heme chaperone-like functions in cells. PMID: 22957700
  48. The ability of C1q to sense both human and bacterial GAPDHs sheds new insights on the role of this important defense collagen molecule in modulating the immune response. PMID: 23086952
  49. SIRT1 functions to retain GAPDH in the cytosol, protecting the enzyme from nuclear translocation via interaction with these two proteins. PMID: 22789853
  50. The present mini review summarizes recent findings relating to the extraglycolytic functions of GAPDH and highlights the significant role this enzyme plays in regulating both cell survival and apoptotic death--{REVIEW} PMID: 21895736

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Subcellular Location Cytoplasm, cytosol, Nucleus, Cytoplasm, perinuclear region, Membrane, Cytoplasm, cytoskeleton
Protein Families Glyceraldehyde-3-phosphate dehydrogenase family
Database Links

HGNC: 4141

OMIM: 138400

KEGG: hsa:2597

STRING: 9606.ENSP00000229239

UniGene: Hs.544577

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