Antibody Validation for Cytoskeletal Proteins

Antibodies against neurofilament-light chain Ab9035 (Western blot), and NFL mouse monoclonal, Cat# MCA-DA2 (immunostaining), tyrosinated α-tubulin (clone YL1/2; ab6160) and stathmin-1 (clone EP1573Y; ab52630) were obtained from Abcam. The specificity of these antibodies and in particular the stathmin antibody has been tested in previous studies [5 (link), 66 (link)]. After stathmin-1 knockout [5 (link)] or lentiviral knockdown [66 (link)] the corresponding Stathmin-1 band was completely abolished in Western blots. Neurofilament-heavy chain antibody (AB5539) was obtained from Millipore, eIF2α (D7D3), p-STAT3^Y705 (D3A7), and Stat3 (124H6) (9139S) antibodies from Cell Signaling Technology. γ-tubulin (clone GTU-88), tau (T-6402), acetylated-α-tubulin (clone 6-11b-1; T7451) and α-tubulin (clone B-5-1-2; T5168) antibodies were purchased from Sigma-Aldrich. The Stathmin 2 (SCG10) antibody was a kind gift from the lab of Dr. Gabriele Grenningloh, EPFL [15 (link)].

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Yadav P., Selvaraj B.T., Bender F.L., Behringer M., Moradi M., Sivadasan R., Dombert B., Blum R., Asan E., Sauer M., Julien J.P, & Sendtner M. (2016). Neurofilament depletion improves microtubule dynamics via modulation of Stat3/stathmin signaling. Acta Neuropathologica, 132, 93-110.

Publication 2016

α-tubulin (clone B-5-1-2; T5168)

Antibodies Antibodies specificity Antibody Clone Heavy chain antibody Mouse Neurofilament Stat3 Stathmin Tubulin Western blot Western blots α tubulin

Corresponding Organization :

Other organizations : University of Würzburg, Institut Universitaire en Santé Mentale de Québec

Top 5 similar protocols

Variable analysis

independent variables

Stathmin-1 knockout [5 (link)]
Lentiviral knockdown of Stathmin-1 [66 (link)]

dependent variables

Stathmin-1 band in Western blots

control variables

Antibodies against neurofilament-light chain Ab9035 (Western blot)
NFL mouse monoclonal, Cat# MCA-DA2 (immunostaining)
Tyrosinated α-tubulin (clone YL1/2; ab6160)
Stathmin-1 (clone EP1573Y; ab52630)
Neurofilament-heavy chain antibody (AB5539)
EIF2α (D7D3)
P-STAT3^Y705 (D3A7)
Stat3 (124H6) (9139S)
γ-tubulin (clone GTU-88)
Tau (T-6402)
Acetylated-α-tubulin (clone 6-11b-1; T7451)
α-tubulin (clone B-5-1-2; T5168)
Stathmin 2 (SCG10) antibody

positive controls

After stathmin-1 knockout [5 (link)] or lentiviral knockdown [66 (link)] the corresponding Stathmin-1 band was completely abolished in Western blots.

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