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GAPDH Monoclonal antibody
GAPDH Monoclonal Antibody for WB, IF/ICC, FC (Intra), IP, ELISA
Host / Isotype
Mouse / IgG2b
Reactivity
human, mouse, rat, pig, zebrafish, yeast, plant and More (7)
Applications
WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA
Conjugate
Unconjugated
11534
CloneNo.
1E6D9
Cat no : 60004-1-Ig
Synonyms
Validation Data Gallery
Tested Applications
| Positive WB detected in | HeLa cells, HepG2 cells, ROS1728 cells, pig brain tissue, zebrafish tissue, whole yeast, whole Nematode tissue, soybean whole plant tissue, arabidopsis whole plant tissue, HEK-293 cells, Jurkat cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells, 4T1 cells, C6 cells, PC-12 cells, C2C12 cells, SP2/0 cells, rat brain tissue, mouse brain tissue |
| Positive IP detected in | HeLa Cells |
| Positive IF/ICC detected in | HeLa cells |
| Positive FC (Intra) detected in | HeLa cells |
Recommended dilution
| Application | Dilution |
|---|---|
| Western Blot (WB) | WB : 1:50000-1:500000 |
| Immunoprecipitation (IP) | IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate |
| Immunofluorescence (IF)/ICC | IF/ICC : 1:400-1:1600 |
| Flow Cytometry (FC) (INTRA) | FC (INTRA) : 0.40 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. | |
Product Information
60004-1-Ig targets GAPDH in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat, pig, zebrafish, yeast, plant samples.
| Tested Reactivity | human, mouse, rat, pig, zebrafish, yeast, plant |
| Cited Reactivity | human, rat, chicken, goat, yeast, deer, whiteflies, tick, treeshrew, cynomorium songaricum |
| Host / Isotype | Mouse / IgG2b |
| Class | Monoclonal |
| 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_2107436 |
| Conjugate | Unconjugated |
| Form | Liquid |
| Purification Method | Protein A 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 -20oC storage. 20ul sizes contain 0.1% BSA. |
Background Information
What is the molecular weight of GAPDH?
The molecular weight of GAPDH is 36 kD.
What is GAPDH?
GAPDH stands for glyceraldehyde-3-phosphate dehydrogenase and is often referred to as a "housekeeping"
protein. It is recognized as an important enzyme involved in metabolic pathways, aiding in the glycolytic
production of ATP and pyruvate (PMID: 21895736).
What is the function of GAPDH?
GAPDH is responsible for catalyzing the phosphorylation of glyceraldehyde-3-phosphate into D-glycerate-1,3-biphosphate. While this function of GAPDH is considered its primary function, GAPDH is also involved, even
critical, in other cell functions, some of which are still being studied. Recent studies have suggested that GAPDH
is a key factor in regulating apoptosis, translocating to the nucleus under certain stressful conditions and acting
as a signaling factor in oxidative stress-induced apoptosis. Other studies indicate that GAPDH plays a role in
transporting vesicles from the endoplasmic reticulum to the Golgi body (PMIDs: 21895736, 12829261, 25859407,
11035021).
Where in the cell is GAPDH localized?
GAPDH is expressed primarily in the cytosolic and membrane regions, with minimal expression in the nucleus
(PMIDs: 21895736, 12829261).
Is there a reason to use an antibody targeting GAPDH if my research is not concerned with glycolysis or nuclear
functions?
Yes! Because GAPDH is often highly expressed throughout different tissues and cell types, GAPDH expression is
useful as a control for Western blots and RT-PCR.
What are the known protein-protein interactions of GAPDH?
GAPDH interacts with other proteins to regulate nuclear translocation and GAPDH catalytic activity. Some of
these proteins include TPPP, SIAH1, CHP1, RILPL1/GOSPEL and FKBP6 (PMIDs: 21895736, 25859407).
Note: For murine tissue samples, conjugated mouse antibody HRP-60004 and rabbit antibody 10494-1-AP are preferable.
Protocols
| Product Specific Protocols | |
|---|---|
| WB protocol for GAPDH antibody 60004-1-Ig | Download protocol |
| IF protocol for GAPDH antibody 60004-1-Ig | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
| Species | Application | Title |
|---|---|---|
Reviews
The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.
FH Sara (Verified Customer) (10-13-2024) | Wonderful antibody!
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FH Rajeshkumar (Verified Customer) (10-10-2024) | It gives very nice and clear blots
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FH pk (Verified Customer) (07-05-2024) | eXCELLENT
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FH S (Verified Customer) (05-16-2024) | Excellent
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FH Alexandra (Verified Customer) (03-07-2024) | Extremely efficient!
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FH Sammy (Verified Customer) (01-20-2024) | Excellent House Keeper. Diluted in 3% BSA PBS Tween (0.2%).
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FH Kamal (Verified Customer) (08-30-2023) | Works well with when diluted in 1X TBST.
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FH Kamal (Verified Customer) (08-30-2023) | Works well with when diluted in 1X TBST.
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FH Parijat (Verified Customer) (08-21-2023) | Works well, no backgrounds
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FH Amy (Verified Customer) (08-07-2023) | Good clean bands for WB of HEK293T cell lysates.
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FH Mohamad (Verified Customer) (07-03-2023) | very good antibody
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FH Angie (Verified Customer) (06-02-2023) | Protein lysates from A549 cells were subjected to SDS-PAGE followed by immunoblotting with 60004-1-Ig (mouse anti-GAPDH antibody) at dilution of 1:80 000 incubated at room temperature for 45 minutes. Secondary antibody: Donkey-anti-mouse (Alexa Fluor 680) at 1:20000 dilution incubated at room temperature for 1 hour .
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FH Udesh (Verified Customer) (09-27-2022) | The antibody was specific and detected a band close to 35 kD in BM-MSCs at 1:5000 dilution, overnight at 4 degrees.
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FH Janelle (Verified Customer) (09-12-2022) | Specific to target
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FH Yuki (Verified Customer) (08-23-2022) | A nice antibody to detect GAPDH in 293T cells
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FH Ralf (Verified Customer) (08-04-2022) | 10µg of total mouse protein were loaded on the gel. Excellent antibodies!
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FH Tsimafei (Verified Customer) (07-15-2022) | 30µg of total protein were loaded on the gel. Antibodies were incubated ON at 4C.
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FH Xin (Verified Customer) (04-15-2022) | A nice antibody to detect GAPDH in both mouse cortex lysates.
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FH Shu Jun (Verified Customer) (04-14-2022) | Not applicable
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FH Weihao (Verified Customer) (04-11-2022) | Great antibody! Clear band.
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FH Charlotte (Verified Customer) (03-09-2022) | Membrane was blocked for 1 hour with 5% nonfat milk in TBS + 0.01% Tween, then probed overnight at 4C with a 1:20,000 dilution of a-GAPDH antibody in blocking buffer. After washing, the membrane was probed with an a-mouse secondary antibody at 1:50,000 in TBST. Under these conditions, the signal is very robust, even at a short exposure (~30 seconds) and has low background. The GAPDH antibody could likely be used at lower concentrations. Quite pleased with how it looks!
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FH Uthra (Verified Customer) (03-03-2022) | Excellent antibody! It gives clean and specific results
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FH Jinwei (Verified Customer) (08-31-2021) | This GAPDH Monoclonal antibody Cat No. 60004-1-Ig works OK.
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FH IMRAN AKHTAR (Verified Customer) (04-15-2021) | This antibody is very good. We have used it many times and with really good results.
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FH Pradeep (Verified Customer) (02-02-2021) | Good antibody for WB.
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FH Yin (Verified Customer) (01-27-2021) | 293T lysate (20 ug) was loaded on 4-12% Bis-Tris gels and transferred to PVDF membrane. Membrane was incubated in 5% BSA in PBS/0.1% Tween-20 for 1h. Membrane was incubated with anti-GAPDH in blocking buffer (1:5000) at 4 degree overnight. After washing, membrane was incubated in anti-mouse-HRP in blocking buffer (1:10000) for 1h at room temperature. Membrane was developed in ECL and imaged on a Biorad chemiluminescence detection system.
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FH Eiko (Verified Customer) (10-16-2020) | This antibody worked quite well in my western blotting. I used 5% skim milk TBS-T for blocking and antibody dilution.
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FH Tom (Verified Customer) (09-28-2020) | HEK293T lysate (10ug) ran on 4-20% gradient gel. Blocking in 5% BSA for 1 hour, followed by HSP27 (1:5000 in block) overnight at 4 degrees. Goat anti-mouse HRP (1:10,000) for 1 hour in block.
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FH Federica (Verified Customer) (06-08-2020) | Excellent antibody.I used in WB on HeLa lysate and the results was extremely clear. No unspecific bands detected. Perfect as loading control.
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FH Alinda (Verified Customer) (01-27-2020) | Good antibody for Western BLOT. antibody diluted in 5%milk.
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FH Aamir (Verified Customer) (01-19-2020) | Good signal for WB
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FH David (Verified Customer) (01-13-2020) | Excellent antibody. Single band at the correct molecular weight. No background and can be used at high dilutions.
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FH saswati (Verified Customer) (01-08-2020) | Need a 1 hr block with 5% milk / TBST as usual.1 hr room temp with the same blocking media with a 1:8000 dilution, followed by secondary anti mouse ( same dilution) , HRP conjugated 1 hr room temp.Detect and document.4 is rating , since a 1:8000 fold is tad bit low, have used better dilution ( 1:100,000) in the past, however that product is not available these days
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FH Emma (Verified Customer) (12-22-2019) | This antibody works well detecting GAPDH in 3t3-L1 and primary mouse embryonic fibroblasts.
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FH Nhu Quynh (Verified Customer) (12-16-2019) | High afffinity and specific antibody. Useful for protein controls in western blots.
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FH Jia (Verified Customer) (11-12-2019) | This antibody works really well.
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FH Juan (Verified Customer) (10-20-2019) | WB, it works well in 1:10000 dilution.
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FH Iram (Verified Customer) (10-01-2019) | Working excellent
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FH RICHA (Verified Customer) (09-25-2019) | Excellent antibody. Clean and specific
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FH Pau (Verified Customer) (09-02-2019) | Excellent and clean Western blots.
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FH Tanusree (Verified Customer) (08-14-2019) | The antibody works good in mouse brain tissue in western blotting.
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FH Praveen (Verified Customer) (04-15-2019) | Good
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FH Shubham (Verified Customer) (04-15-2019) | Very good
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FH Vanessa (Verified Customer) (04-14-2019) | Total cell lysate (5-20 ug) was resolved on 4-12% Bis-Tris gels and transferred to nitrocellulose membrane. Membrane was incubated in blocking buffer (5% BSA in PBS/0.1% Tween-20) for 1h. Membrane was incubated with anti-GAPDH in blocking buffer (1:5000) at 4C overnight. After washing, membrane was incubated in anti-mouse-HRP in blocking bufffer (1:10000) for 2h at room temperature. Membrane was developed in ECF reagent and imaged on a chemiluminescence detection system.
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FH Damaris (Verified Customer) (04-03-2019) | Excellent antibody! It gives clean and specific results
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FH JIMMY (Verified Customer) (03-26-2019) | This antibody offers a good detection of GAPDH on mouse cells such as macrophages and microglia through western blots.
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FH Elisabeth (Verified Customer) (02-05-2019) | Total cell lysate (15 ug) was resolved on 4-12% Bis-Tris gels and transferred to nitrocellulose membrane. Membrane was incubated in blocking buffer (5% milk in PBS/0.1% Tween-20) for 1h. Membrane was incubated with anti-GAPDH in blocking buffer (1:10,000) at 4C overnight. After washing, membrane was incubated in anti-mouse-HRP in blocking bufffer (1:5000) for 2h at room temperature. Membrane was developed in ECL reagent and imaged on a chemiluminescence detection system.
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FH Stephen (Verified Customer) (01-31-2019) | great antibody for WB as a loading control used 1:20000
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FH Kishor (Verified Customer) (01-30-2019) | I working good for Western Blotting (1:5000)
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FH Wei (Verified Customer) (12-04-2018) | In muscle cells, GAPDH antibody shows two bands while in cancer cells there is only one band
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FH Robert (Verified Customer) (12-03-2018) |
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FH Guillaume (Verified Customer) (11-21-2018) | This antibody is useful as a cytoplasmic control for cyto-nuclear extractions.
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FH Ruth (Verified Customer) (11-21-2018) |
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FH Tingting (Verified Customer) (11-20-2018) | The bands of the GADPH looked very good! The antibody used in trophoblast cell and diluted in 1:1000.
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FH marian (Verified Customer) (11-20-2018) |
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FH vanessa (Verified Customer) (11-20-2018) | very efficient product and great customer service
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FH Vermont (Verified Customer) (11-20-2018) | great sensitivity and reproducibility
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FH Saswati (Verified Customer) (11-20-2018) |
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FH Qunfeng (Verified Customer) (01-11-2018) |
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FH Sinu (Verified Customer) (01-10-2018) | Worked well in the western blot
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