Detection of Human, Mouse, and Rat Muscle Phosphofructokinase/PFKM by Western Blot.
Western blot shows lysates of human heart tissue, mouse heart tissue, and rat heart tissue. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti‑Human Muscle Phosphofructokinase/PFKM Monoclonal Antibody (Catalog # MAB7687) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Muscle Phosphofructokinase/PFKM at approximately 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human and Rat Muscle Phosphofructokinase/PFKM by Simple WesternTM.
Simple Western lane view shows lysates of human heart tissue and rat heart tissue, loaded at 0.5 mg/mL. A specific band was detected for Muscle Phosphofructokinase/PFKM at approximately 84 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse/Rat Muscle Phosphofructokinase/PFKM Monoclonal Antibody (Catalog # MAB7687). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Muscle Phosphofructokinase/PFKM
PFKM (PhosphoFructoKinase, Muscle type; also PFK-A and Phosphohexokinase)) is a 85-86 kDa member of the phosphofructokinase family, two domain subfamily of enzymes. It is one of three functionally related enzymes, all expressed by distinct genes. While PFKM is expressed by virtually all cells, its name is based on the fact that skeletal muscle expresses only the PFKM isotype. PFK catalyzes the essentially irreversible phosphorylation of Fru-6-P, forming Fru-1,6-bisP. This determines the rate of cellular glycolysis. As a monomer, PFKM is unstable, and following synthesis, PFKM forms low activity homodimers. High activity comes with tetramerization/oligomerization. In all cases, multimerization is associated with PFKM interaction with various components of the cell cytoskeleton. Human PFKM is 780 amino acids (aa) in length. It contains a 740 aa catalytic region (aa 16-755) plus one utilized phosphorylation site at Thr2. There are at least two potential isoform variants. One contains an alternative start site 71 aa upstream of the standard site, while another shows a deletion of aa 282-312. Proteolysis of PKFM apparently occurs, generating 45-47 kDa fragments. Over aa 674-780, human PFKM shares 98% aa sequence identity with mouse PFKM.
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