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  4. GAPDH Monoclonal Antibody

GAPDH Monoclonal Antibody

  • 货号：
    CSB-MA000071M1m
  • 规格：
    ￥400
  • 图片：

    • 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

  • 其他：

产品详情

  • 产品描述：

    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.

  • 产品名称：
    Mouse anti-Homo sapiens (Human) GAPDH Monoclonal Antibody antibody
  • Uniprot No.：
    P04406
  • 基因名：
  • 别名：
    GAPDH; G3PD; GAPD; MGC88685
  • 宿主：
    Mouse
  • 反应种属：
    Human, Mouse, Rabbit
  • 免疫原：
    Recombinant Human GAPDH protein (2-335AA)
  • 免疫原种属：
    Homo sapiens (Human)
  • 标记方式：
    Non-conjugated
  • 克隆类型：
    Monoclonal Antibody
  • 抗体亚型：
    IgG1
  • 纯化方式：
    >95%, Protein G purified
  • 克隆号：
    10B4E3
  • 浓度：
    It differs from different batches. Please contact us to confirm it.
  • 保存缓冲液：
    Preservative: 0.03% Proclin 300
    Constituents: 50% Glycerol, 0.01M PBS, PH 7.4
  • 产品提供形式：
    Liquid
  • 应用范围：
    ELISA, WB, IHC, IP, IF
  • 推荐稀释比：
    Application Recommended Dilution
    WB 1:5000-1:80000
    IHC 1:200-1:500
    IF 1:50-1:100
    IP 2µl-8µl
  • Protocols：
  • 储存条件：
    Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
  • 货期：
    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.

产品评价

靶点详情

  • 功能：
    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.
  • 基因功能参考文献：
    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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  • 亚细胞定位：
    Cytoplasm, cytosol. Nucleus. Cytoplasm, perinuclear region. Membrane. Cytoplasm, cytoskeleton.
  • 蛋白家族：
    Glyceraldehyde-3-phosphate dehydrogenase family
  • 数据库链接：

    HGNC: 4141

    OMIM: 138400

    KEGG: hsa:2597

    STRING: 9606.ENSP00000229239

    UniGene: Hs.544577