Glycolysis Antibody Kit
Catalog# HAK21008
Glycolysis Antibody Kit
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WB
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IHC-P
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Human
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Mouse
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Rat
General Overview
Kit Components
| Product Includes | Specification | Application | Reactivity | Mw |
|---|---|---|---|---|
| PKM2[HA723370] | 20µl | WB,IF-Cell,IHC-P,FC,IP | Human,Mouse,Rat,Monkey | Predicted band size: 58 kDa |
| GAPDH[ET1601-4] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,FC,IP | Human,Mouse,Rat,Chicken,Drosophila melanogaster | Predicted band size: 36 kDa |
| PDHA1[ET1702-75] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human,Mouse,Rat | Predicted band size: 43 kDa |
| Hexokinase 1[ET1609-28] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human,Mouse,Rat | Predicted band size: 102 kDa |
| Hexokinase II[HA722933] | 20µl | WB,IHC-P,IF-Cell,FC,IP | Human,Mouse,Rat,Monkey | Predicted band size: 102 kDa |
| Lactate Dehydrogenase[ET1608-57] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,FC,IP | Human,Mouse | Predicted band size: 37 kDa |
| PFKP[HA500472] | 20µl | WB,IHC-P | Human,Mouse,Rat | 86 kDa |
| Alpaca anti-Rabbit IgG Fc, Recombinant VHH[HA1031] | 100µl | IP,ELISA,IHC-P,WB | Rabbit |
Product Description
The Glycolysis Antibody Sampler Kit provides an economical means to investigate select enzymes involved in glycolysis. The kit contains enough primary antibody to perform two western blot experiments with each primary antibody.
Product Features
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Background
Glycolysis is the metabolic process by which glucose is converted to pyruvate in a sequence of enzymatic steps. Hexokinase catalyzes the conversion of glucose to glucose-6-phosphate, the first step in glycolysis. Hexokinases I, II, and III are associated with the outer mitochondrial membrane and are critical for maintaining an elevated rate of aerobic glycolysis in cancer cells (Warburg effect). Phosphofructokinase (PFK) catalyzes the phosphorylation of fructose-6-phosphate. Platelet-type phosphofructokinase (PFKP) is expressed in various cell types. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the phosphorylation of glyceraldehyde-3-phosphate. Pyruvate kinase, a glycolytic enzyme, catalyses the conversion of phosphoenolpyruvate to pyruvate. In mammals, the M1 isoform (PKM1) is expressed in most adult tissues. The M2 isoform (PKM2), an alternatively-spliced variant of M1, is expressed during embryonic development. Lactate dehydrogenase (LDH) catalyzes the interconversion of pyruvate and NADH to lactate and NAD+. LDHA expression is induced when the oxygen supply is too low for mitochondrial ATP production. The pyruvate dehydrogenase complex catalyzes the conversion of pyruvate and CoA into acetyl-CoA and CO2 in the presence of NAD+. The reaction of oxidative decarboxylation of pyruvate serves as a critical link between glycolysis and the citric acid cycle and lipid metabolism.
Data Links
Background References
1. WARBURG, O. (1956) Science 123, 309-14.
2. Morrison, N. et al. (1992) Hum Genet 89, 105-6.
3. Barber, R.D. et al. (2005) Physiol Genomics 21, 389-95.
4. Christofk, H.R. et al. (2008) Nature 452, 230-3.
5. Semenza, G.L. et al. (1996) J Biol Chem 271, 32529-37.
6. Semenza, G.L. (2007) Biochem J 405, 1-9.
7. Strumiło, S. (2005) Acta Biochim Pol 52, 759-64.
Images
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All lanes: Western blot analysis of PKM with anti-PKM antibody (R1603-5) at 1:500 dilution.
Lane 1: Wild-type MDA-MB-231 whole cell lysate.
Lane 2: PKM knockout MDA-MB-231 whole cell lysate.
R1603-5 was shown to specifically react with PKM in wild-type MDA-MB-231 cells. No band was observed when PKM knockout sample was tested. Wild-type and PKM knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (R1603-5, 1/500) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Cell lysate was provided by Ubigene Biosciences (Ubigene Biosciences Co., Ltd., Guangzhou, China). -
Western blot analysis of GAPDH on Hela cell lysates with Rabbit anti-GAPDH antibody (ET1601-4).
Hela cell lysates at 10 µg/Lane.
Predicted band size: 36 kDa
Observed band size: 36 kDa
12% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-4) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of Lactate Dehydrogenase on different lysates with Rabbit anti-Lactate Dehydrogenase antibody (ET1608-57) at 1/5,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HEK-293 cell lysate (15 µg/Lane)
Lane 3: A549 cell lysate (15 µg/Lane)
Predicted band size: 37 kDa
Observed band size: 37 kDa
Exposure time: 1 minute 36 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-57) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
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