To assess KCC2 (1:500; #07-432 - Millipore, Temecula, CA; AB_310611) and phospho-Ser940KCC2 (1:1000; p1551-940 - PhosphoSolutions, Aurora, CO; AB_2492213) expression, spinal cord specimens were homogenized with dounce homogenizer (Kontes), followed by 5 passes through a 22 gauge needle in ice-cold buffer, pH 7.5, containing 10 mm Tris, 300 mm sucrose, and a complete mini protease inhibitor mixture (Roche). Crude homogenates were centrifuged at 5000 RCF for 5 min at 4° C. Supernatant was further fractionated at 13,000 RCF for 30 min. After centrifugation, supernatant was collected as cytoplasmic fraction. A membrane rich fraction was then obtained by resuspending the pellet in PBS (50 μl) containing protease inhibitor. All samples were sonicated and stored at −80° C for later processing (Western blotting). N-cadherin (1:1000; #4061 - Cell Signaling, Danvers, MA; AB_2077426) was used to confirm plasma membrane enrichment.
P1551 940
Manufactured by PhosphoSolutions
Sourced in Philippines, United States
The P1551-940 is a laboratory instrument used for the detection and quantification of proteins. It utilizes a specialized technique to measure the level of phosphorylation in protein samples. The core function of this product is to provide researchers with precise data on the phosphorylation status of their samples, which is a fundamental aspect of many biochemical and cell signaling studies.
Automatically generated - may contain errors
Lab products found in correlation
N cadherin, by Cell Signaling Technology (1 mentions)
Complete miniprotease inhibitor mixture, by Roche (1 mentions)
Mab5406, by Merck Group (1 mentions)
Gad67, by Merck Group (1 mentions)
Ab1555p, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
Ab81271, by Abcam (1 mentions)
Calpain 2, by Cell Signaling Technology (1 mentions)
Calpain 1, by Cell Signaling Technology (1 mentions)
Lf sa8001, by AbFrontier (1 mentions)
Lf sa8002, by AbFrontier (1 mentions)
Goat anti mouse igg hrp, by AbFrontier (1 mentions)
2 protocols using p1551 940
1
Quantification of KCC2 and pS940-KCC2
2
Immunoblotting analysis of neuronal proteins
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All commercial antibodies and reagents were purchased from the following sources: NR2A (1:500; AB1555P), NR2B (1:1000; #06-600), KCC2 (1:1000, #07-432), and GAD67 (1:1000, MAB5406) were purchased from Millipore (Billerica, MA, USA). NR2B-phospho S1303 (1:1000; ab81271) and KCC2-NTD (1:1000, ab107452) were purchased from Abcam (Cambridge, MA, USA). Calpain-1 (1:1000, #2556S) and calpain-2 (1:1000; #2539S) were purchased from Cell Signaling Technology (Danvers, MA, USA). KCC2-phospho S940 (1:1000; p1551-940) was purchased from Phosphosolutions (Aurora, CO, USA). β-actin (1:10000, A5316; Sigma-Aldrich, St. Louis, MO, USA) was used as a loading control. Goat anti-rabbit IgG-HRP (1:5000, #LF-SA8002) and goat anti-mouse IgG-HRP (1:5000, #LF-SA8001) were purchased from AbFron-tier Co., Ltd. (Seoul, Korea). N-Methyl-D-aspartic acid (NMDA) (M3262) and MDL-27180 (M6690) were purchased from Sigma-Aldrich.
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