GAPDH Polyclonal antibody

GAPDH Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat, pig, arabidopsis, cabbage, corn, rice and More (7)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, RIP, ELISA, Cell treatment

Conjugate

Unconjugated

Publications

6862

Reviews (11)

Cat no : 10494-1-AP

Synonyms

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Validation Data Gallery

• 

  WB analysis using 10494-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10494-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Raji using 10494-1-AP

  Raji cells were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of K-562 using 10494-1-AP

  K-562 cells were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse heart using 10494-1-AP

  mouse heart tissue were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 10494-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH Antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of arabidopsis whole plant using 10494-1-AP

  arabidopsis whole plant tissue were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of corn whole plant using 10494-1-AP

  corn whole plant tissue were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10494-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 10494-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10494-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa cells using 10494-1-AP

  Western blot of Hela cell with anti-GAPDH (10494-1-AP) at various dilutions.

• 

  WB analysis of PC-13 using 10494-1-AP

  PC-13 cells were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10494-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10494-1-AP (GAPDH antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of A549 using 10494-1-AP

  IP result of anti-GAPDH (IP:10494-1-AP, 3ug; Detection:10494-1-AP 1:3000) with A549 cells lysate 3500ug.

• 

  IHC staining of human breast cancer using 10494-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 10494-1-AP (GAPDH antibody) at dilution of 1:400 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 10494-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 10494-1-AP (GAPDH antibody) at dilution of 1:400 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 10494-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 10494-1-AP (GAPDH antibody) at dilution of 1:400 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 10494-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 10494-1-AP (GAPDH antibody) at dilution of 1:400 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 10494-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer using 10494-1-AP (GAPDH antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human lung cancer using 10494-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer using 10494-1-AP (GAPDH antibody) at dilution of 1:50 (under 40x lens).

• 

  IF Staining of HepG2 using 10494-1-AP

  Immunofluorescent analysis of HepG2 cells, using GAPDH antibody 10494-1-AP at 1:50 dilution and Rhodamine-labeled goat anti-rabbit IgG (red). Blue pseudocolor = DAPI (fluorescent DNA dye).

• 

  IF Staining of HeLa using 10494-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using GAPDH antibody (10494-1-AP) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CoraLite®594 Beta Actin antibody (CL594-66009, Clone: 2D4H5, red).

• 

  FC experiment of HEK-293 using 10494-1-AP

  1x10^6 HEK-293 cells were intracellularly stained with 0.5 ug GAPDH Polyclonal antibody (10494-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.5 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: human placenta tissue, HepG2 cells, HEK-293 cells, HeLa cells, Raji cells, K-562 cells, mouse heart tissue, PC-13 cells, arabidopsis whole plant tissue, corn whole plant tissue, mouse brain tissue, mouse skin tissue, Jurkat cells, NIH/3T3 cells, C6 cells, rat brain tissue, RAW 264.7 cells
• Positive IP detected in: A549 cells
• Positive IHC detected in: human breast cancer tissue, human lung cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells, HepG2 cells
• Positive FC (Intra) detected in: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:40000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.50 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 6570 publications below
• IHC: See 7 publications below
• IF: See 11 publications below
• IP: See 14 publications below
• ELISA: See 2 publications below
• CoIP: See 10 publications below
• ChIP: See 1 publications below
• RIP: See 3 publications below

Product Information

10494-1-AP targets GAPDH in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, RIP, ELISA, Cell treatment applications and shows reactivity with human, mouse, rat, pig, arabidopsis, cabbage, corn, rice samples.

• Tested Reactivity: human, mouse, rat, pig, arabidopsis, cabbage, corn, rice
• Cited Reactivity: human, rat, chicken, goat, sheep, yeast, leech, treeshrew, arabidopsis, zebra finches
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: GAPDH fusion protein Ag0766
• Full Name: glyceraldehyde-3-phosphate dehydrogenase
• Calculated Molecular Weight: 36 kDa
• Observed Molecular Weight: 36 kDa
• GenBank Accession Number: BC004109
• Gene Symbol: GAPDH
• Gene ID (NCBI): 2597
• RRID: AB_2263076
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the phosphorylation of glyceraldehyde-3-phosphate during glycolysis. GAPDH participates in nuclear events including transcription, binding RNA, RNA transportation, DNA replication, DNA repair and apoptosis. Being stably and constitutively expressed at high levels in most tissues and cells, GAPDH is considered a housekeeping protein. It is widely used as a control for RT-PCR and also loading control in electrophoresis and Western blotting. GAPDH is normally expressed in cellular cytoplasm or membrane, but can occasionally translocate to the nucleus after the addition of post-translational modifications such as S-nitrosylation. This antibody is raised against full length GAPDH of human origin. It can recognize the 36 kDa GAPDH protein in most cells/tissues. In addition, a band below 36 kDa can always be detected as the isoform or spliced product of GAPDH (PMID: 23885286, 23877755, 19368702). Please note that some physiological factors, such as hypoxia and diabetes, increase GAPDH expression in certain cell types.

Publications

Species	Application	Title
human	WB	Nature; 7-Dehydrocholesterol dictates ferroptosis sensitivity; Authors - Yaxu Li
human		Cell; TEX264 drives selective autophagy of DNA lesions to promote DNA repair and cell survival; Authors - Pauline Lascaux
human	WB	Cell Res; Mitochondria-localized cGAS suppresses ferroptosis to promote cancer progression; Authors - Shiqiao Qiu
mouse	WB	Nature; MAP3K2-regulated intestinal stromal cells define a distinct stem cell niche.; Authors - Ningbo Wu
human	WB	Nature; Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts tropism and fusogenicity.; Authors - Bo Meng
human,mouse	WB	Nature; Feeding induces cholesterol biosynthesis via the mTORC1-USP20-HMGCR axis.; Authors - Xiao-Yi Lu

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Parisa (Verified Customer) (09-26-2023)

High sensitivy and specificity. We were using a anti mouse-GAPDH antibody and this anti rabbit-GAPDH antibody gave us more flexibility for probing the blot.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Hek293

GAPDH Antibody Western Blot validation (1:1000 dilution) in Hek293 (Cat no:10494-1-AP)

FH

Sathyanarayani (Verified Customer) (05-12-2023)

This GADPH loading control antibody produced strong bands at 36kDa and I would definitely recommend using it for western blots.

• Applications: Western Blot
• Cell Tissue Type: Mouse lysed blood

FH

Verena (Verified Customer) (11-09-2022)

Gives a nice, strong and specific signal in Western Blot.

• Applications: Western Blot
• Primary Antibody Dilution: 1:5000
• Cell Tissue Type: Mouse

FH

Christine (Verified Customer) (10-27-2022)

Very good loading control antibody, strong signal when used at 1:15,000 over night, can probably be diluted much more.

• Applications: Western Blot
• Primary Antibody Dilution: 1:15,000
• Cell Tissue Type: HEK293T

FH

Janelle (Verified Customer) (09-12-2022)

Perfect for WB!

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000

FH

Jake (Verified Customer) (11-30-2021)

GAPDH Ab works perfectly on western blot at low dilutions, will definitely buy again.

• Applications: Western Blot
• Primary Antibody Dilution: 1:10000
• Cell Tissue Type: BV-2 Cell Line

FH

Meg (Verified Customer) (02-25-2020)

It worked well for western blotting.

• Applications: Western Blot,
• Primary Antibody Dilution: 1:3000
• Cell Tissue Type: THP-1

FH

Yuan (Verified Customer) (10-15-2019)

GAPDH is working very well on seminal plasma and blood plasma.

• Applications: Western Blot,
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: Human seminal plasma and blood plasma

FH

Hailey (Verified Customer) (10-07-2019)

This antibody is a fantastic loading control! I use this as a loading control for all my western blots where protein loading needs to be precise.

• Applications: Western Blot,
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: Mouse testis

GAPDH Antibody Western Blot, validation (1:500 dilution) in Mouse testis (Cat no:10494-1-AP)

FH

Prasanth (Verified Customer) (08-19-2019)

The band is so clear, and very specific band with almost no background noise for GAPDH

• Applications: Western Blot,
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Mouse Fibroblasts

FH

Shannon (Verified Customer) (12-11-2018)

• Applications: Cell culture,
• Cell Tissue Type: rodent

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