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8337
Glycolysis Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Glycolysis Antibody Sampler Kit #8337

Citations (36)

Simple Western™ analysis of lysates (0.1 mg/mL) from Cos-7 cells using Hexokinase II (C64G5) Rabbit mAb #2867. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Flow cytometric analysis of Caki-1 cells using LDHA (C4B5) Rabbit mAb (solid line) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using GAPDH (D16H11) XP ® Rabbit mAb #5174. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from various cell types using Hexokinase I (C35C4) Rabbit mAb.
Western blot analysis of various cell extracts using Hexokinase II (C64G5) Rabbit mAb (upper) and β-Actin (13E5) Rabbit mAb #4970 (lower).
Western blot analysis of extracts from various cell types using PKM1/2 (C103A3) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Pyruvate Dehydrogenase (C54G1) Rabbit mAb.
Western blot analysis of extracts from various cell types using LDHA (C4B5) Rabbit mAb.
Western blot analysis of extracts from various cell lines and mouse skeletal muscle using PKM2 (D78A4) XP® Rabbit mAb (upper) or GAPDH (14C10) Rabbit mAb #2118.
Western blot analysis of extracts from various cell lines using GAPDH (D16H11) XP® Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from various cell lines using PFKP (D4B2) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Hexokinase I (C35C4) Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells using PKM1/2 (C103A3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Western blot analysis of various cell lines using Pyruvate Dehydrogenase (C54G1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using LDHA (C4B5) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using PKM2 (D78A4) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GAPDH (D16H11) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Hexokinase I (C35C4) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Pyruvate Dehydrogenase (C54G1) Rabbit mAb.
Confocal immunofluorescent analysis of MCF-7 cells using LDHA (C4B5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human lymphoma using PKM2 (D78A4) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse colon using GAPDH (D16H11) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells using Hexokinase I (C35C4) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Pyruvate Dehydrogenase (C54G1) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human skeletal muscle using PKM2 (D78A4) XP® Rabbit mAb. Note the lack of staining in the skeletal muscle cells which do not express PKM2 while vessels within the tissue stain positively.
Confocal immunofluorescent analysis of C2C12 cells using GAPDH (D16H11) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of fixed frozen mouse colon labeled with Hexokinase I (C35C4) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182 (right, red), and DAPI #4083 (right, blue).
Confocal immunofluorescent analysis of A204 cells using PKM2 (D78A4) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of fixed frozen mouse pancreas labeled with Hexokinase I (C35C4) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182 (right, red), and DAPI #4083 (right, blue).
Flow cytometric analysis of 293T cells, transfected with PKM2 siRNA (blue) or mock transfected (green), using PKM2 (D78A4) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.
Confocal immunofluorescent analysis of fixed frozen mouse kidney labeled with Hexokinase I (C35C4) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182 (right, red), and DAPI #4083 (right, blue).
To Purchase # 8337
Cat. # Size Qty. Price
8337T
1 Kit  (8 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
PKM2 (D78A4) XP® Rabbit mAb 4053 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk 60 Rabbit IgG
GAPDH (D16H11) XP® Rabbit mAb 5174 20 µl
  • WB
  • IHC
  • IF
H M R Mk 37 Rabbit IgG
Pyruvate Dehydrogenase (C54G1) Rabbit mAb 3205 20 µl
  • WB
  • IHC
H M R Mk 43 Rabbit IgG
Hexokinase I (C35C4) Rabbit mAb 2024 20 µl
  • WB
  • IP
  • IHC
  • IF
H M 102 Rabbit IgG
Hexokinase II (C64G5) Rabbit mAb 2867 20 µl
  • WB
H M R Mk 102 Rabbit 
LDHA (C4B5) Rabbit mAb 3582 20 µl
  • WB
  • IHC
  • IF
  • F
H Mk 37 Rabbit IgG
PKM1/2 (C103A3) Rabbit mAb 3190 20 µl
  • WB
  • IF
H M R Mk 60 Rabbit IgG
PFKP (D4B2) Rabbit mAb 8164 20 µl
  • WB
  • IP
H Mk 80 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

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.

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).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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