Western Blot Analysis of Brain Proteins

Brain tissues were homogenized and lysed in SDS-Lysis buffer and western blotting was performed using a protocol described in our previous work (Singh et al., 2012 (link)). Samples (50 μg protein) were blotted using antibodies against GATA1 (ab28839, Abcam; 1:500 in 5% BSA), NRXN2 (ABN97, Milipore; 1:400 in 5% BSA), CAMK2A (Prestige Antibody, Sigma, 1:300 in 5% BSA), and β-ACTIN (sc47778, SantaCruz; 1:1000 in 5% BSA).

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Mitra S., Sameer Kumar G.S., Jyothi Lakshmi B., Thakur S, & Kumar S. (2018). Absence of Wdr13 Gene Predisposes Mice to Mild Social Isolation – Chronic Stress, Leading to Depression-Like Phenotype Associated With Differential Expression of Synaptic Proteins. Frontiers in Molecular Neuroscience, 11, 133.

Publication 2018

ab28839 Prestige Antibody sc47778

Actin Antibodies Antibody Brain Buffer Gata1 Prestige Protein Tissues

Corresponding Organization : Centre for Cellular and Molecular Biology

Other organizations : Vimta (India)

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Variable analysis

independent variables

Homogenization and lysis of brain tissues in SDS-Lysis buffer

dependent variables

Protein expression levels of GATA1, NRXN2, CAMK2A, and β-ACTIN

control variables

Protein loading (50 μg per sample)
Antibody dilutions (GATA1: 1:500, NRXN2: 1:400, CAMK2A: 1:300, β-ACTIN: 1:1000)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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