重组
RabMAb

重组Anti-GAPDH抗体[EPR16891] - Loading Control (ab181602)

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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

      

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    see all

  • 别名

    • 38 kDa BFA-dependent ADP-ribosylation substrate antibody
    • aging associated gene 9 protein antibody
    • Aging-associated gene 9 protein antibody
    • BARS-38 antibody
    • cb609 antibody
    • EC 1.2.1.12 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-PDH antibody
    • Glyceraldehyde-3-phosphate dehydrogenase antibody
    • HEL-S-162eP antibody
    • KNC-NDS6 antibody
    • MGC102544 antibody
    • MGC102546 antibody
    • MGC103190 antibody
    • MGC103191 antibody
    • MGC105239 antibody
    • MGC127711 antibody
    • MGC88685 antibody
    • OCAS, p38 component antibody
    • OCT1 coactivator in S phase, 38-KD component antibody
    • peptidyl cysteine S nitrosylase GAPDH antibody
    • Peptidyl-cysteine S-nitrosylase GAPDH antibody
    • wu:fb33a10 antibody
    see all

图片

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    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 - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    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.

  • Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    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 Cytometry - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Flow Cytometry - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    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.

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    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.

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    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.

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    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

发表研究结果有使用 ab181602？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab181602 被引用在 461 文献中.

  • Gao Z  et al. Regulatory effects of lncRNA ATB targeting miR-200c on proliferation and apoptosis of colorectal cancer cells. J Cell Biochem 121:332-343 (2020). PubMed: 31222825
  • Chen JQ  et al. MicroRNA-212-3p inhibits the Proliferation and Invasion of Human Hepatocellular Carcinoma Cells by Suppressing CTGF expression. Sci Rep 9:9820 (2019). PubMed: 31285444
  • Shan L  et al. High glucose promotes annulus fibrosus cell apoptosis through activating the JNK and p38 MAPK pathways. Biosci Rep 39:N/A (2019). PubMed: 31221817
  • Zhang B  et al. Urate transport capacity of glucose transporter 9 and urate transporter 1 in cartilage chondrocytes. Mol Med Rep 20:1645-1654 (2019). PubMed: 31257523
  • Shang L  et al. MicroRNA-148a-3p promotes survival and migration of endothelial cells isolated from Apoe deficient mice through restricting circular RNA 0003575. Gene 711:143948 (2019). PubMed: 31255737
View all Publications for this product

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1-10 of 19 Abreviews or Q&A

Western blot abreview for Anti-GAPDH antibody [EPR16891] - Loading Control

 Good
Abreviews
abreview image
Application
Western blot
Sample
Chinese hamster Cell lysate - whole cell (Chinese Hamster Ovary (CHO))
Gel Running Conditions
Non-reduced Denaturing (12% SDS-GEL)
Loading amount
50 µg
Specification
Chinese Hamster Ovary (CHO)
Blocking step
Western Breeze kit as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0% · Temperature: 21°C
Read More

Pavlos Kotidis

Verified customer

提交于 Mar 23 2020

Western blot abreview for Anti-GAPDH antibody [EPR16891] - Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
10 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Jan 27 2020

Western blot abreview for Anti-GAPDH antibody [EPR16891] - Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (L Cells (ATCC® CRL-2648))
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
20 µg
Specification
L Cells (ATCC® CRL-2648)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Jan 20 2020

Western blot abreview for Anti-GAPDH antibody [EPR16891]

 Good
Abreviews
abreview image
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
Read More

Dr. Haibo Wang

Verified customer

提交于 Feb 27 2018

Western blot abreview for Anti-GAPDH antibody [EPR16891]

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Nov 29 2016

Western blot abreview for Anti-GAPDH antibody [EPR16891]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Oct 25 2016

Western blot abreview for Anti-GAPDH antibody [EPR16891]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Oct 25 2016

Western blot abreview for Anti-GAPDH antibody [EPR16891]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Oct 25 2016

Western blot abreview for Anti-GAPDH antibody [EPR16891]

 Excellent
Abreviews
abreview image
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
Read More

Maral Tajerian

Verified customer

提交于 Jul 15 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [EPR16891]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Feb 22 2016

1-10 of 19 Abreviews or Q&A

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