Name: Anti-GAPDH antibody [6C5] - Loading Control
SKU: ab8245
Rating: 5 (89 reviews)

产品名称

Anti-GAPDH抗体[6C5] - Loading Control
参阅全部 GAPDH 一抗
描述

小鼠单克隆抗体[6C5] to GAPDH - Loading Control
宿主

Mouse
特异性

This antibody can be used as a loading control antibody, however we also recommend ab9484 as an alternative. GAPDH is a 146 kDa tetramer composed of four 30-40 kDa subunits. There is no cross-reaction with GAPDH from yeast. Preliminary data indicates that the antibody recognizes the monomer (36 kDa) and also the dimer forms of GAPDH, but not the tetrameric form of the protein.
经测试应用

适用于: ELISA, ICC, WB, ICC/IF, IHC-Frmore details
种属反应性

与反应: Mouse, Rat, Rabbit, Chicken, Hamster, Cat, Dog, Human, Pig, Xenopus laevis, Fish, Monkey, Zebrafish, Baboon, African green monkey
预测可用于: Horse, Guinea pig, Xenopus tropicalis不与反应: Goat, Cow, Saccharomyces cerevisiae
免疫原

Rabbit muscle GAPDH.
阳性对照
- ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells
常规说明

Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor^® 488 (ab150113).
See other anti-mouse secondary antibodies that can be used with this antibody.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
存储溶液

pH: 7.40
Preservative: 0.09% Sodium azide
Constituent: PBS
Concentration information loading...
纯度

Protein A purified
纯化说明

Chromatography on protein A Sepharose
克隆

单克隆
克隆编号

6C5
骨髓瘤

Sp2/0
同种型

IgG1
研究领域

Our Abpromise guarantee covers the use of ab8245 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
ELISA		Use at an assay dependent concentration.
ICC		Use at an assay dependent concentration.
WB		1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa). PubMed: 16450009
ICC/IF		Use a concentration of 1 - 5 µg/ml.
IHC-Fr		1/500.

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.
Target information above from: UniProt accession P04406 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 493876 Cat
- Entrez Gene: 374193 Chicken
- Entrez Gene: 403755 Dog
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 396823 Pig
- Entrez Gene: 100009074 Rabbit
- Entrez Gene: 24383 Rat
- Entrez Gene: 685186 Rat
- Entrez Gene: 380259 Xenopus laevis
- Entrez Gene: 317743 Zebrafish
- Omim: 138400 Human
- SwissProt: Q9N2D5 Cat
- SwissProt: P00356 Chicken
- SwissProt: Q28259 Dog
- SwissProt: P04406 Human
- SwissProt: P16858 Mouse
- SwissProt: P00355 Pig
- SwissProt: P46406 Rabbit
- SwissProt: P04797 Rat
- SwissProt: P51469 Xenopus laevis
- SwissProt: Q5XJ10 Zebrafish
- Unigene: 544577 Human
- Unigene: 592355 Human
- Unigene: 598320 Human
- Unigene: 304088 Mouse
- Unigene: 309092 Mouse
- Unigene: 317779 Mouse
- Unigene: 343110 Mouse
- Unigene: 392463 Mouse
- Unigene: 392480 Mouse
- Unigene: 414470 Mouse
- Unigene: 458138 Mouse
- Unigene: 458416 Mouse
- Unigene: 475698 Mouse
- Unigene: 129558 Rat
- Unigene: 91450 Rat
- Unigene: 995 Xenopus laevis
- Unigene: 35640 Zebrafish
see all
别名
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- G3PDH antibody
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- GAPDH antibody
- Glyceraldehyde 3 phosphate dehydrogenase antibody
- Glyceraldehyde-3-phosphate dehydrogenase antibody
- HEL-S-162eP antibody
- KNC-NDS6 antibody
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- MGC102546 antibody
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- MGC105239 antibody
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- OCAS, p38 component antibody
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see all

Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

All lanes : Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 2.5 µg/ml

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) Nuclear
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Alexa Fluor anti-mouse at 1/5000 dilution

Performed under reducing conditions.

Predicted band size: 40.2 kDa
Observed band size: 37 kDa
why is the actual band size different from the predicted?

Fluorescence detection of secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

ab8245 staining GAPDH in SV40LT-SMC cells.

The cells were fixed with 4% formaldehyde (10 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)This image is courtesy of an anonymous Abreview

All lanes : Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

Lane 1 : Mouse hippocampus whole cell lysate
Lane 2 : Rat hippocampus whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP-conjugated Rabbit anti-mouse at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 40.2 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

ab8245 staining GAPDH in NIH/3T3 (Mouse embryo fibroblast cell line) cells.

The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 1 μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

ab8245 staining GAPDH in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5 μg/ml and ab6046 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) at 2 μg/ml (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.

Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)This image is courtesy of an anonymous Abreview

All lanes : Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 1/20000 dilution

Lane 1 : Rabbit aorta, whole tissue lysate at 10 µg
Lane 2 : Blank
Lane 3 : Mouse aorta, whole tissue lysate at 5 µg
Lane 4 : Mouse aorta, whole tissue lysate at 7.5 µg
Lane 5 : Mouse aorta, whole tissue lysate at 10 µg
Lane 6 : Mouse aorta, whole tissue lysate at 12.5 µg
Lane 7 : Mouse aorta, whole tissue lysate at 15 µg

Secondary
All lanes : HRP conjugated sheep anti-mouse antibody at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 40.2 kDa

Exposure time: 30 seconds
See Abreview
Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 10 µg/ml + Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Predicted band size: 40.2 kDa

Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Zhao W et al. Long noncoding RNA NSCLCAT1 increases non-small cell lung cancer cell invasion and migration through the Hippo signaling pathway by interacting with CDH1. FASEB J 33:1151-1166 (2019). Read more (PubMed: 30148675) »
Yuan J et al. GLIS2 redundancy causes chemoresistance and poor prognosis of gastric cancer based on co-expression network analysis. Oncol Rep 41:191-201 (2019). Read more (PubMed: 30320360) »

See all 1311 Publications for this product

发表研究结果有使用 ab8245？请让我们知道，以便我们可以引用本数据表中的参考文章。

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Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Mouse Cell lysate - other (MEF (cytosol fraction))

Gel Running Conditions

Reduced Denaturing (12)

Loading amount

10 µg

Specification

MEF (cytosol fraction)

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Jan 09 2019

Application

Western blot

Sample

Human Cell lysate - whole cell (H1299 cells)

Gel Running Conditions

Reduced Denaturing (12)

Loading amount

25 µg

Specification

H1299 cells

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 Sep 21 2018

Application

Western blot

Sample

Mouse Tissue lysate - whole (Brain, heart, liver)

Gel Running Conditions

Reduced Denaturing (12% gel)

Loading amount

25 µg

Specification

Brain, heart, liver

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 Sep 21 2018

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - other (A549)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

50 µg

Specification

A549

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 May 22 2018

Application

Western blot

Sample

Rat Tissue lysate - whole (rat hippocampus tissue)

Gel Running Conditions

Non-reduced Denaturing

Loading amount

50 µg

Treatment

RIPA buffer

Specification

rat hippocampus tissue

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Sep 14 2017

Application

Western blot

Sample

Bos taurus Tissue lysate - whole (Lane 1Bovine Liver and Lane 2 Bovine mammary gland)

Gel Running Conditions

Reduced Denaturing (4-15% Gradient)

Loading amount

20 µg

Specification

Lane 1Bovine Liver and Lane 2 Bovine mammary gland

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Julie Kim

Verified customer

提交于 May 12 2017

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Mouse Tissue lysate - whole (Mouse whole brain)

Gel Running Conditions

Reduced Denaturing

Loading amount

20 µg

Specification

Mouse whole brain

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

提交于 May 04 2017

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Rat Tissue lysate - whole (Liver)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

50 µg

Specification

Liver

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

提交于 Feb 24 2017

Abcam has not validated the combination of species/application used in this Abreview.

Application

Western blot

Sample

Caenorhabditis elegans Cell lysate - whole cell (whole lysates)

Gel Running Conditions

Non-reduced Denaturing

Loading amount

50 µg

Specification

whole lysates

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Joey Wu

Verified customer

提交于 Jan 25 2017

Application

Western blot

Sample

Ferret Tissue lysate - whole (Ferret whole brain tissue)

Gel Running Conditions

Reduced Denaturing (3-8% Tris-acetate gel, NuPage)

Loading amount

40 µg

Specification

Ferret whole brain tissue

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Francis Djankpa

Verified customer

提交于 Jul 11 2016

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定制方案及合作关系

Anti-GAPDH抗体[6C5] - Loading Control (ab8245)

概述

性能

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应用

靶标

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实验方案

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文献

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客户评价及客户问答

Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

Western blot abreview for Anti-GAPDH antibody [6C5]

Western blot abreview for Anti-GAPDH antibody [6C5]

Western blot abreview for Anti-GAPDH antibody [6C5]

Western blot abreview for Anti-GAPDH antibody [6C5]

Western blot abreview for Anti-GAPDH antibody [6C5]

Western blot abreview for Anti-GAPDH antibody [6C5]

Western blot abreview for Anti-GAPDH antibody [6C5]