Product Pathways - Glucose Metabolism
Hexokinase I (C35C4) Rabbit mAb (Alexa Fluor® 647 Conjugate) #3540
PhosphoSitePlus® protein, site, and accession data: HK1
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| IF-IC F | H M | Endogenous | Rabbit IgG |
Applications Key:
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 3540:
- Flow, Immunofluorescence*
* Product-specific protocol.
Specificity / Sensitivity
Hexokinase I (C35C4) Rabbit mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of total hexokinase I protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of human hexokinase I. This antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.
Flow Cytometry
Flow cytometric analysis of MCF-7 cells using Hexokinase I (C35C4) Rabbit mAb (Alexa Fluor® 647 Conjugate) (green) compared to XP® Rabbit (DA1E) mAb IgG Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (red).
IF-IC
Confocal immunofluorescent analysis of MCF-7 cells using Hexokinase I (C35C4) Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue pseudocolor) and MitoTracker® Red CMXRos (red) showing colocalization with mitochondria. Nuclei have been labeled with p53 (1C12) Mouse mAb (Alexa Fluor® 488 Conjugate) #2015 (green).
Description
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Hexokinase I (C35C4) Rabbit mAb #2024.
Background
Hexokinase catalyzes the conversion of glucose to glucose-6-phosphate, the first step in glycolysis. Four distinct mammalian hexokinase isoforms, designated as hexokinase I, II, III and IV (glucokinase), have been identified. 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) in order to compensate for the increased energy demands associated with rapid cell growth and proliferation (2,3).
- Warburg, O. (1956) Science 123, 309-314.
- Semenza, G.L. (2000) Crit. Rev. Biochem. Mol. Biol. 35, 71-103.
- Smith, T.A. (2000) Br. J. Biomed. Sci. 57, 170-178.
Application References
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Companion Products
- 2024 Hexokinase I (C35C4) Rabbit mAb
- 3689 Hexokinase I (C35C4) Rabbit mAb (Alexa Fluor® 488 Conjugate)
- 2985 Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate)
Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 from Epitomics, Inc.
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
