Name: Anti-GAPDH antibody [6C5] - Loading Control
SKU: ab8245
Rating: 5 (94 reviews)

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Key features and details

Mouse monoclonal [6C5] to GAPDH - Loading Control
Suitable for: WB, ICC/IF
Reacts with: Mouse, Rat, Rabbit, Human
Isotype: IgG1

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一次抗体

タンパク質

二次抗体

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一次抗体 - Mouse Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
WB, ICC/IF

タンパク質 - Recombinant Mouse GAPDH protein (ab202148)
SDS-PAGE, MS

二次抗体 - Goat Anti-Mouse IgG H&L (HRP) (ab205719)
WB, IP, ELISA, IHC-P

関連製品

製品名

Anti-GAPDH antibody [6C5] - Loading Control
GAPDH 一次抗体製品一覧
製品の詳細

Mouse monoclonal [6C5] to GAPDH - Loading Control
由来種

Mouse
特異性

This GAPDH antibody can be used as a loading control antibody. 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 GAPDH antibody- loading control ab8245 recognizes the monomer (36 kDa) and also the dimer forms of GAPDH, but not the tetrameric form of the protein.
アプリケーション

適用あり: WB, ICC/IFmore details
種交差性

交差種: Mouse, Rat, Rabbit, Human
交差が予測される動物種: Horse, Chicken, Guinea pig, Hamster, Cat, Dog, Pig, Xenopus laevis, Fish, Monkey, Zebrafish, Baboon, African green monkey, Xenopus tropicalis非交差種: Goat, Cow, Saccharomyces cerevisiae
免疫原

Rabbit muscle GAPDH.
ポジティブ･コントロール
- ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells

製品の状態

Liquid
保存方法

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.
バッファー

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

Protein A purified
特記事項（精製）

Chromatography on protein A Sepharose
ポリ/モノ

モノクローナル
クローン名

6C5
ミエローマ

Sp2/0
アイソタイプ

IgG1
研究分野

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.

アプリケーション	Abreviews	特記事項
WB		1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa).
ICC/IF		Use a concentration of 1 - 5 µg/ml.

機能

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.
パスウェイ

Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
配列類似性

Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
翻訳後修飾

S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated.
細胞内局在

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
参照データベース
- 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
別名
- 38 kDa BFA-dependent ADP-ribosylation substrate antibody
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- 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

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 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)

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
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 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).
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
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.

Immunohistochemistry protocols

Click here to view the general protocols

ab8245 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab8245 は 2205 報の論文で使用されています。

He B et al. MicroRNA-326 decreases tau phosphorylation and neuron apoptosis through inhibition of the JNK signaling pathway by targeting VAV1 in Alzheimer's disease. J Cell Physiol 235:480-493 (2020). PubMed: 31385301
Sun H et al. miR-4429 sensitized cervical cancer cells to irradiation by targeting RAD51. J Cell Physiol 235:185-193 (2020). PubMed: 31190335
Li L et al. XIST/miR-376c-5p/OPN axis modulates the influence of proinflammatory M1 macrophages on osteoarthritis chondrocyte apoptosis. J Cell Physiol 235:281-293 (2020). PubMed: 31215024
Xu J & Zhang J LncRNA TP73-AS1 is a novel regulator in cervical cancer via miR-329-3p/ARF1 axis. J Cell Biochem 121:344-352 (2020). PubMed: 31232491
Doccini S et al. Proteomic and functional analyses in disease models reveal CLN5 protein involvement in mitochondrial dysfunction. Cell Death Discov 6:18 (2020). WB ; Human . PubMed: 32257390

View all Publications for this product

レビューと Q&A

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1-10 of 104 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (U87 cells)

Gel Running Conditions

Non-reduced Non-Denaturing (Native) (4-20% gel)

Loading amount

15 µg

Treatment

bevacizumab 2mg/ml for 24hrs, 48hrs

Specification

U87 cells

Blocking step

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

Abcam user community

Verified customer

投稿 May 11 2020

Application

Western blot

Sample

Rat Cell lysate - whole cell (Hippocampal culture)

Gel Running Conditions

Reduced Denaturing (4-15% gradient)

Loading amount

20 µg

Specification

Hippocampal culture

Blocking step

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

Abcam user community

Verified customer

投稿 Jan 31 2020

Application

Western blot

Sample

Mouse Tissue lysate - whole (Cerebellum)

Gel Running Conditions

Reduced Denaturing (4-15% gradient)

Loading amount

20 µg

Specification

Cerebellum

Blocking step

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

Abcam user community

Verified customer

投稿 Jan 31 2020

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

Abcam user community

Verified customer

投稿 Jan 31 2020

Application

Western blot

Sample

Rat Tissue lysate - whole (Brain tissue)

Gel Running Conditions

Reduced Denaturing (8% SDS Running gel)

Loading amount

80 µg

Treatment

10mg ketamine

Specification

Brain tissue

Blocking step

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

Collin Gagne

Verified customer

投稿 Nov 12 2019

Application

Western blot

Sample

Human Tissue lysate - whole (skin tissue)

Gel Running Conditions

Reduced Denaturing (10% gel)

Loading amount

30 µg

Treatment

UV B (100mJ/cm2)

Specification

skin tissue

Blocking step

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

Abcam user community

Verified customer

投稿 Apr 08 2019

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

投稿 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

投稿 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

投稿 Sep 21 2018

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

投稿 May 22 2018

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Key features and details

製品の概要

製品名

製品の詳細

由来種

特異性

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

製品の特性

製品の状態

保存方法

バッファー

精製度

特記事項（精製）

ポリ/モノ

クローン名

ミエローマ

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Related Products

アプリケーション

ターゲット情報

機能

パスウェイ

配列類似性

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

参考文献

レビューと Q&A

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] - 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] - 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]