概述

  • 产品名称

    Anti-GAPDH抗体[EPR16891] - Loading Control
    参阅全部 GAPDH 一抗
  • 描述

    兔单克隆抗体[EPR16891] to GAPDH - Loading Control
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • 种属反应性

    与反应: Mouse, Rat, Rabbit, Chicken, Human, Fish, Monkey, Zebrafish, Xenopus tropicalis
  • 免疫原

    Recombinant fragment within Mouse GAPDH aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: P16858

  • 阳性对照

    • WB: HeLa, UMNSAH/DF-1, Jurkat, COS-1, RAW 264.7 and PC-12 whole cell lysates; Human fetal brain and heart lysates; Xenopus(X. tropicalis) muscle lysate; Zebrafish lysate; Mouse kidney and spleen lysates; Rat brain lysate. IHC-P: Human transitional cell carcinoma of bladder, Mouse spleen and Rat spleen tissues. ICC/IF: HeLa cells. Flow: Jurkat cells. IP: HeLa whole cell extract
  • 常规说明

      

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab181602 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
Flow Cyt 1/180.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/60.

靶标

  • 功能

    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 (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.
  • 通路

    Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
  • 序列相似性

    Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
  • 翻译后修饰

    S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
    ISGylated.
  • 细胞定位

    Cytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
  • Information by UniProt
  • 数据库链接

  • 别名

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    • HEL-S-162eP antibody
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图片

  • All lanes : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution

    Lane 1 : Mouse kidney lysates
    Lane 2 : Mouse spleen lysates
    Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 5 : Rat brain lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling GAPDH with ab181602 at dilution of 1/500. The secondary antibody used was Alexa Fluor® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with ab150120 (AlexaFluor®594 goat anti-mouse secondary, 1/500). The negative controls are shown in the bottom middle and right hand panels- for negative control 1 primary antibody (ab181602; 1/500) and secondary antibody (ab150120; 1/500) was used. For negative control 2 primary antibody (ab7291; 1/500) and secondary antibody (ab150077; 1/400) was used.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • GAPDH was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181602 at 1/60 dilution. Western blot was performed from the immunoprecipitate using ab181602 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract.

    Lane 2: PBS instead of HeLa whole cell extract.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181602 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • All lanes : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/50000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : Xenopus(X. tropicalis) muscle lysates
    Lane 3 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of human bladder is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution

    Lane 1 : COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates
    Lane 2 : Zebrafish lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution

    Lane 1 : Human fetal brain lysates
    Lane 2 : Human fetal heart lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

文献

This product has been referenced in:

See all 383 Publications for this product

客户评价及客户问答

1-10 of 16 Abreviews or Q&A

Application
Western blot
Sample
Escherichia coli Cell lysate - whole cell (E. coli)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
10 µg
Specification
E. coli
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Haibo Wang

Verified customer

提交于 Feb 27 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Fibroblasts)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
Fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

提交于 Nov 29 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain, liver, retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Brain, liver, retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Oct 25 2016

Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Brain, retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Brain, retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Oct 25 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (ARPE19, hfRPE, HEK293)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
ARPE19, hfRPE, HEK293
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Oct 25 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (skin)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
skin
Blocking step
Commercial blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 23°C

Maral Tajerian

Verified customer

提交于 Jul 15 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (undifferentiated Human ES cells on MEFs)
Permeabilization
Yes - see blocking buffer
Specification
undifferentiated Human ES cells on MEFs
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Feb 22 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (HUES7 ES cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
HUES7 ES cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

提交于 Feb 04 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEPG2)
Specification
HEPG2
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Nov 18 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (HEPG2)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
20 µg
Specification
HEPG2
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

提交于 Nov 13 2015

1-10 of 16 Abreviews or Q&A

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