Whole brain lysates were harvested using 20 mM HEPES, 320 mM sucrose with protease inhibitors (0.1 µg/µl Leupeptin/Aprotinin and 1 mM PMSF). Protein concentration of lysate was determined using the Bio-Rad protein concentration assay and equal amounts of protein were loaded and subjected to SDS-PAGE gel electrophoresis followed by transfer to PVDF membranes (Immobilon-P). Membranes were probed with mouse anti-Kirrel3 1:100 (Neuromab Clone N321C/49; catalog no. 75–333) that detects the long and short isoforms of Kirrel3, rat anti-HSC70/HSP73 1:10,000 (Enzo Life Sciences), or rabbit anti-β-actin 1:1,000 (Cell Signaling Technology), followed with appropriate HRP-conjugated secondary antibodies. Blots were developed using SuperSignal West Femto Kit (Thermo Fisher Scientific). Quantification of relative protein levels was performed on scanned immunoblots using Fiji software.
Rat anti hsc70 hsp73
Manufactured by Enzo Life Sciences
The Rat anti-HSC70/HSP73 is a monoclonal antibody that recognizes the heat shock cognate 70 kDa protein (HSC70), also known as heat shock protein 73 (HSP73). HSC70 is a constitutively expressed member of the HSP70 family of molecular chaperones, which play a crucial role in protein folding, transport, and degradation processes within the cell.
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2 protocols using rat anti hsc70 hsp73
1
Quantifying Kirrel3 Protein Levels
2
Quantifying Kirrel3 Protein Levels
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Whole brain lysates were harvested using 20 mM HEPES, 320 mM sucrose with protease inhibitors (0.1 µg/µl Leupeptin/Aprotinin and 1 mM PMSF). Protein concentration of lysate was determined using the Bio-Rad protein concentration assay and equal amounts of protein were loaded and subjected to SDS-PAGE gel electrophoresis followed by transfer to PVDF membranes (Immobilon-P). Membranes were probed with mouse anti-Kirrel3 1:100 (Neuromab Clone N321C/49; catalog no. 75–333) that detects the long and short isoforms of Kirrel3, rat anti-HSC70/HSP73 1:10,000 (Enzo Life Sciences), or rabbit anti-β-actin 1:1,000 (Cell Signaling Technology), followed with appropriate HRP-conjugated secondary antibodies. Blots were developed using SuperSignal West Femto Kit (Thermo Fisher Scientific). Quantification of relative protein levels was performed on scanned immunoblots using Fiji software.
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