Name: Anti-GAPDH antibody [EPR16891] - Loading Control
Brand: RabMAb
SKU: ab181602
Rating: 5 (22 reviews)

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16891] to GAPDH - Loading Control
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Chicken, Human, Zebrafish, African green monkey, Xenopus tropicalis

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free FITC HRP PE

Product name

Anti-GAPDH antibody [EPR16891] - Loading Control
See all GAPDH primary antibodies
Description

Rabbit monoclonal [EPR16891] to GAPDH - Loading Control
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Chicken, Human, Zebrafish, African green monkey, Xenopus tropicalis
Predicted to work with: Rabbit, Fish
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- 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
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16891
Isotype

IgG
Research areas

The Abpromise guarantee

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.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/180. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB	(19)	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	(3)	1/500.
IP		1/60.

Notes
Flow Cyt (Intra) 1/180. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.
IP 1/60.

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: 374193 Chicken
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 100009074 Rabbit
- Entrez Gene: 24383 Rat
- Entrez Gene: 685186 Rat
- Entrez Gene: 317743 Zebrafish
- Omim: 138400 Human
- SwissProt: P00356 Chicken
- SwissProt: P04406 Human
- SwissProt: P16858 Mouse
- SwissProt: P46406 Rabbit
- SwissProt: P04797 Rat
- 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: 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 [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 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)

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)

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

Intracellular 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)

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

Different batches of ab181602 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 36 kDa.
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)

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)

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)

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

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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Publishing research using ab181602? Please let us know so that we can cite the reference in this datasheet.

ab181602 has been referenced in 1571 publications.

Yan Y et al. The circular RNA hsa_circ_0001394 promotes hepatocellular carcinoma progression by targeting the miR-527/UBE2A axis. Cell Death Discov 8:81 (2022). WB ; Human . PubMed: 35210429
Liu X et al. m6A methylated EphA2 and VEGFA through IGF2BP2/3 regulation promotes vasculogenic mimicry in colorectal cancer via PI3K/AKT and ERK1/2 signaling. Cell Death Dis 13:483 (2022). WB ; Human . PubMed: 35595748
Liu F et al. LncRNA LIFR-AS1 overexpression suppressed the progression of serous ovarian carcinoma. J Clin Lab Anal N/A:e25470 (2022). WB ; Human . PubMed: 35778954
Guo S et al. Ligustilide alleviates the insulin resistance, lipid accumulation, and pathological injury with elevated phosphorylated AMPK level in rats with diabetes mellitus. J Recept Signal Transduct Res 41:85-92 (2021). PubMed: 32643505
Lu W et al. Reprogramming immunosuppressive myeloid cells facilitates immunotherapy for colorectal cancer. EMBO Mol Med 13:e12798 (2021). PubMed: 33283987

View all Publications for this product

Customer reviews and Q&As

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1-10 of 22 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (MCF7 cells lysate)

Gel Running Conditions

Reduced Denaturing (4–20% precast polyacrylamide gel)

Loading amount

10 µg

Specification

MCF7 cells lysate

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 16 2022

Application

Western blot

Sample

Human Cell lysate - whole cell (SH-SY5Y)

Gel Running Conditions

Reduced Denaturing (4-12% Bris-Tris)

Loading amount

75000 cells

Specification

SH-SY5Y

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%

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 16 2022

Application

Western blot

Sample

Human Cell lysate - whole cell (Lung cancer cells)

Gel Running Conditions

Reduced Denaturing (10-14)

Loading amount

40 µg

Specification

Lung cancer cells

Blocking step

BSA 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.

MISS. Marie Klimontova

Verified customer

Submitted Apr 09 2021

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

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.

Pavlos Kotidis

Verified customer

Submitted Mar 23 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 27 2020

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

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 20 2020

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

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. Haibo Wang

Verified customer

Submitted Feb 27 2018

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

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 29 2016

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

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 25 2016

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

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 25 2016

1-10 of 22 Abreviews or Q&A

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

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

Certificate of Compliance

References (1571)

Customer reviews and Q&As

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

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

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

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

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

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

Western blot abreview for Anti-GAPDH antibody [EPR16891]

Western blot abreview for Anti-GAPDH antibody [EPR16891]

Western blot abreview for Anti-GAPDH antibody [EPR16891]

Western blot abreview for Anti-GAPDH antibody [EPR16891]

Post-translational
modifications