GAPDH Monoclonal Antibody

Datasheet

Code	CSB-MA000071M2m
Size	US$200
Image	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, MG-63 whole cell lysate, U251 whole cell lysate, SH-SY5Y 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: 5min Western Blot Positive WB detected in: SP2/0 whole cell lysate, NIH/3T3 whole cell lysate, Raw264.7 whole cell lysate, Mouse heart tissue, Mouse kidney 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 Western Blot Positive WB detected in: Rat heart tissue, Rat liver tissue, Rat spleen tissue, Rat brain tissue, Rat skeletal tissue, Rat stomach tissue, Rat kidney 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 Western Blot Positive WB detected in: Rabbit heart tissue, Rabbit kidney 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 Western Blot Positive WB detected in: Hela whole cell lysate at 10μg, 5μg, 2.5μg, 1.25μg, 0.625μg, 0.3125μg, 0.15625μg, 0.078μg 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: 15μg hela whole cell lysate GAPDH antibody at 1:10000, 1:20000, 1:40000, 1:80000, 1:160000 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-MA000071M2m diluted at 1:500 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 Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. IHC image of CSB-MA000071M2m diluted at 1:500 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 Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. IHC image of CSB-MA000071M2m diluted at 1:500 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 Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Immunofluorescence staining of Hela cells with (CSB-MA000071M2m)at 1:50, 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-MA000071M2m)at 1:50, 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-MA000071M2m in Hela whole cell lysate. Lane 2: CSB-MA000071M2m (5µl) + Hela whole cell lysate (500µg) Lane 3: Hela whole cell lysate (10µg) For western blotting, the blot was detected with CSB-MA000071M2m at 1:5000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:2000
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Product Details
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Target Background

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, Mouse

Immunogen

Recombinant Human GAPDH protein (2-335AA)

Immunogen Species

Human

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

IgG1

Clone No.

10H4D6

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

Recommended Dilution

Application	Recommended Dilution
WB	1:5000-1:160000
IHC	1:200-1:500
IF	1:50-1:100
IP	2µl-8µl

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunoprecipitation (IP) Protocol
Immunofluorescence (IF) 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.

Customer Reviews and Q&A

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Target Background

Function	Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. 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. 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. 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. Also plays a role in innate immunity by promoting TNF-induced NF-kappa-B activation and type I interferon production, via interaction with TRAF2 and TRAF3, 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.
Gene References into Functions	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 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 Nitric oxide-induced GAPDH aggregation specifically induces mitochondrial dysfunction via permeability transition pore opening, leading to cell death. PMID: 28167533 GAPDH may act as a chaperone in heme transfer to downstream areas PMID: 28315300 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 Monoclonal Antibodies DSHB-hGAPDH-2G7 and DSHB-hGAPDH-4B7 Against Human Glyceraldehyde-3-Phosphate Dehydrogenase. PMID: 27556912 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 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 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 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 This review will summarize our current understanding of GAPDH-mediated regulation of RNA function PMID: 26564736 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 In 60 % of patients with type 2 diabetes, a reversible inhibition of GAPDH is observed. PMID: 25189828 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 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 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 Genetic variants in GAPDH confer susceptibility to sporadic Parkinson's disease in a Chinese Han population. PMID: 26258539 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 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 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 suggests that GAPDH aggregates accelerate Abeta amyloidogenesis, subsequently leading to mitochondrial dysfunction and neuronal cell death in the pathogenesis of AD PMID: 26359500 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 The activity of GAPDS was significantly positively correlated with sperm motility and negatively with the incidence of infertility. PMID: 26255202 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 deregulated GAPDH expression promotes NF-kappaB-dependent induction of HIF-1alpha and has a key role in lymphoma vascularization and aggressiveness PMID: 25394713 analysis of how flux through GAPDH is a limiting step in aerobic glycolysis PMID: 25009227 astrocytic production of D-serine is modulated by glycolytic activity via interactions between GAPDH and SRR. PMID: 25870284 Dimer and tetramer interface residues in adenine-uridine rich elements are important for GAPDH-RNA binding. PMID: 25451934 Siah1 is a substrate of ASK1 for activation of the GAPDH-Siah1 oxidative stress signaling cascade. PMID: 25391652 GAPDH expression is deregulated during melanoma progression. PMID: 25550585 Oxidation of an exposed methionine instigates the aggregation of glyceraldehyde-3-phosphate dehydrogenase. PMID: 25086035 MZF-1 binds to and positively regulates the GAPDH promoter, indicating a role for GAPDH in calcitriol-mediated signaling. PMID: 25065746 The protein encoded by this gene contains a peptide that displays antimicrobial activity against E. coli, P. aeruginosa, and C. albicans. PMID: 22832495 GAPDH gene over expression in resected tumor samples is an adverse prognostic factor in non small cell lung cancer. PMID: 23988223 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 TG2-dependent GAPDH deamidation was suggested to participate in actin cytoskeletal remodeling. PMID: 24375405 acetylation of GAPDH (K254) is reversibly regulated by the acetyltransferase PCAF and the deacetylase HDAC5. PMID: 24362262 GAPDH binds to active Akt, leading to Bcl-xL increase and escape from caspase-independent cell death. PMID: 23645209 GAPDH is a moonlighting protein that functions as a glycolytic enzyme as well as a uracil DNA glycosylase. PMID: 20727968 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 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 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 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 NleB, a bacterial glycosyltransferase, targets GAPDH function to inhibit NF-kappaB activation. PMID: 23332158 GAPDH binds to alkylated, single-stranded, double-stranded and telomeric sequences in a drug-dependent and DNA sequence/structure-dependent manner. PMID: 23409959 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 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 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 SIRT1 functions to retain GAPDH in the cytosol, protecting the enzyme from nuclear translocation via interaction with these two proteins. PMID: 22789853 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 Show More Hide All
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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GAPDH Monoclonal Antibody

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