Name: Anti-GAPDH antibody [6C5] - Loading Control
SKU: ab8245
Rating: 5 (100 reviews)

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

Key features and details

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

Product name

Anti-GAPDH antibody [6C5] - Loading Control
See all GAPDH primary antibodies
Description

Mouse monoclonal [6C5] to GAPDH - Loading Control
Host species

Mouse
Specificity

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.
Tested applications

Suitable for: WB, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Predicted to work with: Horse, Chicken, Guinea pig, Hamster, Cat, Dog, Pig, Xenopus laevis, Fish, Monkey, Zebrafish, Baboon, Xenopus tropicalisDoes not react with: Goat, Cow, Saccharomyces cerevisiae
Immunogen

Full length native protein (purified) corresponding to GAPDH.
Database link: P46406
Positive control
- ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells. WB: HeLa, A431, Jurkat, HEK-293, Raji whole cell lysate.
General notes

This product switched from ascites to tissue culture supernatant on 31 July 2017. Lot numbers higher than [GR291713] will be from tissue culture supernatant.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Form

Liquid
Storage instructions

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.
Storage buffer

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

Protein A purified
Purification notes

Chromatography on protein A Sepharose
Clonality

Monoclonal
Clone number

6C5
Myeloma

Sp2/0
Isotype

IgG1
Research areas

The Abpromise guarantee

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.

Application	Abreviews	Notes
WB	(96)	1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa).
ICC/IF	(1)	Use a concentration of 1 - 5 µg/ml.

Notes
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.

Function

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.
Pathway

Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
Sequence similarities

Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
Post-translational
modifications

S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated.
Cellular localization

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
Database links
- 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: 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: 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
Alternative names
- 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

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 (Rat SV40-transfected aorta smooth cell line) 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).
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
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab8245? Please let us know so that we can cite the reference in this datasheet.

ab8245 has been referenced in 3490 publications.

Chen X et al. KLF4 downregulates FGF21 to activate inflammatory injury and oxidative stress of LPS-induced ATDC5 cells via SIRT1/NF-?B/p53 signaling. Mol Med Rep 25:N/A (2022). WB ; Human . PubMed: 35293599
Wang H et al. KLF4 regulates TERT expression in alveolar epithelial cells in pulmonary fibrosis. Cell Death Dis 13:435 (2022). WB ; Human . PubMed: 35508454
Hu Y et al. Curcumol enhances cisplatin sensitivity of gastric cancer: involvement of microRNA-7 and the nuclear factor-kappa B/snail family transcriptional repressor 1 axis. Bioengineered 13:11668-11683 (2022). WB ; Human . PubMed: 35510522
Zhang H et al. m6A methyltransferase METTL3-induced lncRNA SNHG17 promotes lung adenocarcinoma gefitinib resistance by epigenetically repressing LATS2 expression. Cell Death Dis 13:657 (2022). WB ; Human . PubMed: 35902569
Cheng A et al. ATM loss disrupts the autophagy-lysosomal pathway. Autophagy 17:1998-2010 (2021). PubMed: 32757690

View all Publications for this product

Customer reviews and Q&As

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1-10 of 110 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (T-ALL cell line)

Gel Running Conditions

Reduced Denaturing (12)

Loading amount

34 µg

Specification

T-ALL cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 09 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (Cell line)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

69 µg

Specification

Cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 28 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Cell line)

Gel Running Conditions

Reduced Denaturing (gradient gel)

Loading amount

61 µg

Specification

Cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 13 2021

Application

Western blot

Sample

Mouse Cell lysate - nuclear (Cell line)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

36 µg

Treatment

Untreated and treated cells have been loaded on the gel and GAPDH has been used as loading control.

Specification

Cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 07 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (Hela cells)

Gel Running Conditions

Non-reduced Denaturing (4-12% BIS-TRIS gradient, MOPS running buffer)

Loading amount

20 µg

Specification

Hela cells

Blocking step

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 22 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (hek293)

Gel Running Conditions

Reduced Denaturing (4-12%)

Loading amount

25 µg

Specification

hek293

Blocking step

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Robert Kupp

Verified customer

Submitted Oct 19 2020

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted 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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted 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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted 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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 31 2020

1-10 of 110 Abreviews or Q&A

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Key features and details

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Purification notes

Clonality

Clone number

Myeloma

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Pathway

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (3490)

Customer reviews and Q&As

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] - Loading Control

Post-translational
modifications