Anti-GAPDH抗体- Loading Control (ab9483)

概述

  • 产品名称
    Anti-GAPDH抗体- Loading Control
    参阅全部 GAPDH 一抗
  • 描述
    山羊多克隆抗体to GAPDH - Loading Control
  • 经测试应用
    适用于: WB, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Full length native protein from human erythrocytes.

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; NIH3T3. This antibody also gave a positive signal in Human brain tissue lysate.

性能

应用

Our Abpromise guarantee covers the use of ab9483 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 1/1000. Detects a band of approximately 37 kDa (predicted molecular weight: 35.8 kDa).
ICC/IF Use a concentration of 1 µg/ml.

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
    • Aging associated gene 9 protein antibody
    • Epididymis secretory sperm binding protein Li 162eP antibody
    • G3P_HUMAN antibody
    • G3PD antibody
    • G3PDH antibody
    • GAPD antibody
    • GAPDH antibody
    • Glyceraldehyde 3 phosphate dehydrogenase antibody
    • Glyceraldehyde-3-phosphate dehydrogenase antibody
    • HEL S 162eP antibody
    • Peptidyl-cysteine S-nitrosylase GAPDH antibody
    see all

Anti-GAPDH antibody - Loading Control 图像

  • All lanes : Anti-GAPDH antibody - Loading Control (ab9483) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Rabbit polyclonal to Goat IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 35.8 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab9483 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9483, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-goat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 1µg/ml.
  • ab9483 staining GAPDH in HeLa by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehydeand. Samples were incubated with primary antibody (1/200) for 30 minutes. An undiluted Alexa Fluor® 568-conjugated Donkey anti-goat polyclonal was used as the secondary antibody.

    See Abreview

Anti-GAPDH antibody - Loading Control (ab9483)参考文献

This product has been referenced in:
  • Foley B  et al. Editor's Highlight: Screening ToxCast Prioritized Chemicals for PPARG Function in a Human Adipose-Derived Stem Cell Model of Adipogenesis. Toxicol Sci 155:85-100 (2017). Human . Read more (PubMed: 27664422) »
  • Wei D  et al. Genistein suppresses retinoblastoma cell viability and growth and induces apoptosis by upregulating miR-145 and inhibiting its target ABCE1. Mol Vis 23:385-394 (2017). WB . Read more (PubMed: 28706438) »

See all 65 Publications for this product

Product Wall

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (U2OS osteosarcoma)
Specification
U2OS osteosarcoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Feb 21 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (skeletal muscle)
Loading amount
50 µg
Specification
skeletal muscle
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Sep 25 2012

Thank you very much for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial from a new lot on the order *** which should arrive tomorrow.

Please keep me ud...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (aorta)
Loading amount
20 µg
Specification
aorta
Gel Running Conditions
Reduced Denaturing (Bis-tris 4-12%)
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 May 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Loading amount
50 µg
Specification
U2OS cells
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycine)
Blocking step
normal horse serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Natalie Farny

Verified customer

提交于 Oct 23 2009

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Username

Dr. Sophie Pezet

Verified customer

提交于 Jul 31 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

提交于 Oct 03 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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