GAPDH Recombinant antibody

GAPDH Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, ELISA

Conjugate

Unconjugated

Publications

3

CloneNo.

1H18

Cat no : 81640-5-RR

Synonyms

1H18

---

Validation Data Gallery

• 

  WB analysis using 81640-5-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 81640-5-RR (GAPDH antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  IF Staining of HeLa using 81640-5-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using GAPDH antibody (81640-5-RR, Clone: 1H18 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

• 

  Affinity and Kinetic Characterization of 81640-5-RR

  Biolayer interferometry (BLl) kinetic assays of 81640-5-RR against Human GAPDH were performed. The affinity constant is 10.4 nM.

---

Tested Applications

• Positive WB detected in: mouse brain tissue, rat brain tissue, NIH/3T3 cells, HSC-T6 cells, K-562 cells, Jurakat cells, HEK-293 cells, HeLa cells
• Positive IF/ICC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000

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 3 publications below

Product Information

81640-5-RR targets GAPDH in WB, IF/ICC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• 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
• 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 -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	Eur J Pharmacol; Tumoral EIF4EBP1 regulates the crosstalk between tumor-associated macrophages and tumor cells in MRTK; Authors - Xin Wu
	WB	Mol Carcinog; Potentially functional genetic variants in interferon regulatory factor family genes are associated with colorectal cancer survival; Authors - Xiaoxia Tong
rat	WB	Front Immunol; Astrocyte-derived exosomal miR-378a-5p mitigates cerebral ischemic neuroinflammation by modulating NLRP3-mediated pyroptosis; Authors - Ruiting Sun