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Glycolysis Antibody Sampler Kit #8337

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
PKM2 (D78A4) XP® Rabbit mAb #4053	40 µl	W IP IHC-P IF-IC	H M R Mk	60	Rabbit IgG
GAPDH (D16H11) XP® Rabbit mAb #5174	40 µl	W IHC-P IF-IC	H M R Mk (Pg)	37	Rabbit
Pyruvate Dehydrogenase (C54G1) Rabbit mAb #3205	40 µl	W IHC-P	H M R Mk	43	Rabbit IgG
Hexokinase I (C35C4) Rabbit mAb #2024	40 µl	W IP IHC-P IF-IC	H M	102	Rabbit IgG
Hexokinase II (C64G5) Rabbit mAb #2867	40 µl	W IHC-P IF-IC	H M R Mk	102	Rabbit IgG
LDHA (C4B5) Rabbit mAb #3582	40 µl	W IHC-P IF-IC	H Mk	37	Rabbit IgG
PKM1/2 (C103A3) Rabbit mAb #3190	40 µl	W IF-IC	H M R Mk	60	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat
PFKP (D4B2) Rabbit mAb #8164	40 µl	W IP	H Mk	80	Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

GAPDH (D16H11) XP® Rabbit mAb, Hexokinase I (C35C4) Rabbit mAb, Hexokinase II (C64G5) Rabbit mAb, LDHA (C4B5) Rabbit mAb, PKM2 (D78A4) XP® Rabbit mAb, and PFKP (D4B2) Rabbit mAb all detect endogenous levels of their respective total proteins. Pyruvate Dehydrogenase (C54G1) Rabbit mAb detects endogenous levels of total Pyruvate Dehydrogenase α1 subunit. PKM1/2 (C103A3) Rabbit mAb detects endogenous levels of total PKM (including M1 and M2) protein.

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 four western blot experiments with each primary antibody.

Source / Purification

Monoclonal antibodies were produced by immunizing animals with a synthetic peptide corresponding to the respective sequences of human GAPDH, Hexokinase I, Hexokinase II, LDHA, PKM2, Pyruvate Dehydrogenase or PFKP protein.

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) (1). Phosphofructokinase (PFK) catalyzes the phosphorylation of fructose-6-phosphate. Platelet-type phosphofructokinase (PFKP) is expressed in various cell types (2). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the phosphorylation of glyceraldehyde-3-phosphate (3). 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 (4). Lactate dehydrogenase (LDH) catalyzes the interconversion of pyruvate and NADH to lactate and NAD+ (5). LDHA expression is induced when the oxygen supply is too low for mitochondrial ATP production (6). 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 (7).

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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

2024:

IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

2867:

IHC / Paraffin, Immunofluorescence, Western Blotting

3190:

Immunofluorescence, Western Blotting

3205:

IHC / Paraffin, Western Blotting

3582:

IHC / Paraffin, Immunofluorescence, Western Blotting

4053:

IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

5174:

IHC / Paraffin, Immunofluorescence, Western Blotting

8164:

Immunoprecipitation, Western Blotting

Companion Products

• 7071 Phototope^®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9998 BSA
• 9999 Nonfat Dry Milk
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
• 7722 Blue Loading Buffer Pack
• 7723 Red Loading Buffer Pack
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7075 Anti-biotin, HRP-linked Antibody
• 9803 Cell Lysis Buffer (10X)

Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063) from Epitomics, Inc.

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For Research Use Only. Not For Use In Diagnostic Procedures.