Test GAPDH Monoclonal Antibody

CSB-MA000071M1mTest

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Size:

50μl100μl

US$120

Quantity:

Species Reactivity: Human, Mouse, Rabbit

Raised in: Mouse

Application: ELISA, WB, IHC, IP, IF

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Product Details
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Target Background

Product Details

Alternative Names

GAPDH; G3PD; GAPD; MGC88685

Target Names

GAPDH

Raised in

Mouse

Uniprot NO.

P04406

Species Reactivity

Human, Mouse, Rabbit

Immunogen

Recombinant Human GAPDH protein (2-335AA)

Immunogen Species

Human

Conjugate

Non-conjugated

Isotype

IgG1

Clone No.

10B4E3

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, PH 7.4

Concentration

It differs from different batches. Please contact us to confirm it.

Form

Liquid

Tested Applications

ELISA, WB, IHC, IP, IF

Recommended Dilution

Application	Recommended Dilution
WB	1:5000-1:80000
IHC	1:200-1:500
IF	1:50-1:100
IP	2µ

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.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Protocols

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

Datasheet & COA

Antibody FAQs

Images

Western Blot
Positive WB detected in: U87 whole cell lysate, PC-3 whole cell lysate, 293 whole cell lysate, U251 whole cell lysate, A549 whole cell lysate, A375 whole cell lysate, MG-63 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: 10s
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 lung 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: 10s
Western Blot
Positive WB detected in: Rabbit heart tissue, Rabbit kidney tissue, Rabbit 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: 10s
Western Blot
Positive WB detected in: Hela whole cell lysate at 10μg, 5μg, 2.5μg, 1.25μg, 0.625μg 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: 10s
Western Blot
Positive WB detected in: 15μg hela whole cell lysate GAPDH antibody at 1:10000, 1:20000, 1:40000, 1:80000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 36 KDa
Observed band size: 36 KDa
Exposure time: 10s
IHC image of CSB-MA000071M1m 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-MA000071M1m 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-MA000071M1m 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-MA000071M1m)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-MA000071M1m)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-MA000071M1m in Hela whole cell lysate. Lane 2: CSB-MA000071M1m (5µl) + Hela whole cell lysate (500µg)
Lane 3: Hela whole cell lysate (10µg)
For western blotting, the blot was detected with CSB-MA000071M1m at 1:5000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:2000

Description

This GAPDH monoclonal antibody was raised by fusion of B lymphocytes with immortal cell cultures to produce hybridomas (A Recombinant Human GAPDH protein was used in the immunization process). Hybridomas would produce many copies of GAPDH monoclonal antibody. The specificity of this GAPDH monoclonal antibody makes it extremely efficient for binding of antigen within a mixture of GAPDH. In addition, this antibody has been validated in ELISA, WB, IHC, IP, IF.

GAPDH (G3PD) is the abbreviation of glyceraldehyde-3-phosphate dehydrogenase, which is an enzyme in glycolysis and consists of 4 subunits of 30-40 kDa. The molecular weight is 146 kDa. The enzyme gene is a house-keeping gene, which is expressed at a high level in almost all tissues. The protein expression level in the same cell or tissue is generally constant and is not induced by the partial recognition sites contained. The influence of the substance remains constant, so it is widely used as a standardized internal reference for the extraction of total RNA, poly(A)+ RNA, Western blot and other experimental operations.
GAPDH is generally believed to mainly exert its glycolytic activity in the cytoplasmic matrix, but the latest research has shown that it also has a location on cells and biofilms. For example, when cells are stimulated, the content of GAPDH in the nucleus is significantly increased. When stimulated by neurotoxins, GAPDH first accumulates in the Golgi apparatus, and then enters the nucleus from the Golgi apparatus. The content of GAPDH in the nucleus of Xinhecheng GAPDH also increased significantly during oxidative stress. In addition, GAPDH is often seen on the biomembrane during membrane fusion and vesicle transport.

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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