Quantifying Astrocyte Protein Expression

Cultured human ONH astrocytes were lysed with lysis buffer as described previously (Noh et al., 2013 (link)). The primary antibodies included rabbit polyclonal anti-NR1 and NR2A antibody (1:1000; Cell Signaling Technology, Inc., Danvers, MA), rabbit polyclonal anti-GLAST (EAAT1, 1:500; Santa Cruz Biotechnology Inc., Santa Cruz, CA), mouse monoclonal anti-DRP1 antibody (1:1000; BD Transduction Laboratories), rabbit polyclonal anti-phospho-DRP1 at Ser616 antibody (1:1000; Cell Signaling) and mouse monoclonal anti-actin antibody (1:5000; Millipore). The scanned film images were analyzed with ImageJ (National Institute of Health, MD, USA) and band densities were normalized to the band densities for actin.

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Ju W.K., Kim K.Y., Noh Y.H., Hoshijima M., Lukas T.J., Ellisman M.H., Weinreb R.N, & Perkins G.A. (2014). Increased Mitochondrial Fission and Volume Density by Blocking Glutamate Excitotoxicity Protect Glaucomatous Optic Nerve Head Astrocytes. Glia, 63(5), 736-753.

Publication 2014

rabbit polyclonal anti-NR1 and NR2A antibody mouse monoclonal anti-DRP1 antibody rabbit polyclonal anti-phospho-DRP1 at Ser616 antibody mouse monoclonal anti-actin antibody

Actin Anti antibody Antibodies Antibody Astrocytes Buffer Human Monoclonal antibody Mouse Nr2a Rabbit

Corresponding Organization : University of California, San Diego

Other organizations : Northwestern University

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Variable analysis

independent variables

Lysis of cultured human ONH astrocytes with lysis buffer

dependent variables

Protein expression levels of NR1
Protein expression levels of NR2A
Protein expression levels of GLAST (EAAT1)
Protein expression levels of DRP1
Phosphorylation levels of DRP1 at Ser616

control variables

Actin protein expression levels (for normalization)

controls

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

Annotations

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