Cells transfected with Pkm siRNA were harvested 4 d after transfection. Preparation of whole-cell extracts and immunoblotting were done as described.25 (link) Antibodies used were against Ser473 phosphorylated AKT (Cell Signaling Technologies, 4058), GAPDH (Abcam, Ab8245) and PKM (Sigma-Aldrich, SAB4200095).
Sab4200095
Manufactured by Merck Group
The SAB4200095 is a laboratory equipment product manufactured by Merck Group. It is a specialized instrument designed for scientific research and analysis purposes. The core function of this product is to perform specific laboratory tasks, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
2 protocols using sab4200095
1
Immunoblotting of PKM Silenced Cells
2
Sciatic Nerve Protein Extraction and Western Blot
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Sciatic nerves were dissected from mice, after removal of the epineurium and perineurium, the nerves were frozen in liquid nitrogen and conserved at -80°C. Protein were extracted from whole sciatic nerves after homogenization by sonication in standard RIPA lysis buffer and the protein concentration was measured using a BCA protein assay kit (Pierce). 20 μg proteins were directly analyzed by western blot using standard procedures with 12% SDS–PAGE and transferred on PVDF membranes for immunoblotting. Primary antibodies: Rabbit anti-Pkm1 1/2500 (NBP2-14833, Novus Biologicals); Rabbit anti-Pkm2 1/2500 (SAB4200095, Sigma Aldrich); Mouse anti-α-β-Actin 1/10000 (C1.AC-15, #A1978, Sigma Aldrich). Secondary antibodies, Peroxidase Goat anti Rabbit or Mouse (H+L) (Jackson Immuno Research) were used at 1/10000 dilution.
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