Glycolysis Antibody Kit
概述
试剂盒组分
产品包括 | 数量 | 应用 | 反应性 | MW(kDa) |
---|---|---|---|---|
PKM2[R1603-5] | 20µl | WB,IF-Cell,IHC-P,FC | Human,Mouse,Rat | 58 kDa |
GAPDH[ET1601-4] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,FC,IP | Human,Mouse,Rat,Chicken,Zebrafish | Predicted band size: 36 kDa |
PDHA1[ET1702-75] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human,Mouse,Rat | 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[HA500186] | 20µl | WB,IF-Cell,IHC-P,FC | Human,Mouse,Rat | Predicted band size: 102 kDa |
Lactate Dehydrogenase[ET1608-57] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,FC,IP | Human,Mouse,Rat,Zebrafish | Predicted band size: 37 kDa |
PFKP[HA500472] | 20µl | WB,IHC-P | Human,Mouse,Rat | 86 kDa |
HRP Conjugated Alpaca anti-Rabbit IgG FC, Recombinant VHH[HA1031] | 100µl | IP,ELISA,IHC-P,WB | Rabbit |
产品描述
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.
产品特性
存储缓冲液
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
背景
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.
数据链接
SwissProt: P52480 Mouse
SwissProt: P11980 Rat
SwissProt: P04406 Human
SwissProt: P16858 Mouse
SwissProt: P04797 Rat
SwissProt: P08559 Human
SwissProt: P35486 Mouse
SwissProt: P26284 Rat
SwissProt: P19367 Human
SwissProt: P17710 Mouse
SwissProt: P05708 Rat
SwissProt: P52789 Human
SwissProt: O08528 Mouse
SwissProt: P27881 Rat
SwissProt: P07195 Human
SwissProt: P00338 Human
SwissProt: P16125 Mouse
SwissProt: P06151 Mouse
SwissProt: P42123 Rat
SwissProt: P04642 Rat
SwissProt: Q01813 Human
SwissProt: Q9WUA3 Mouse
SwissProt: P47860 Rat
背景文献
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.
图片
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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.