Detecting Recombinant chIL-7 in Cells

The recombinant chIL-7 in the culture medium was detected with Western blot as previously described (Wen et al., 2013 (link)) using Anti-Myc mAb (Santa Cruz Biotechnologies). For the intracellular chIL-7 detection, the transfected cells were lysed with lysis buffer (5 mM Tris-HCl, 25 mM KCl, 2 mM EGTA, 2 mM EDTA, 1% NP-40, 150 mM NaCl and protease inhibitors) and then subjected to Western blot described as above.

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Huo S., Wang L., Zhang Y., Zhang J., Zuo Y., Xu J., Cui D., Li X, & Zhong F. (2016). Molecular cloning of chicken IL-7 and characterization of its antiviral activity against IBDV in vivo. Poultry Science, 95(11), 2647-2654.

Publication 2016

Anti-Myc mAb

Buffer Cells Culture medium Edta Egta Intracellular Nacl Np 40 Protease inhibitors Tris Western blot

Corresponding Organization : Yanshan University

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Variable analysis

independent variables

Recombinant chIL-7 in the culture medium

dependent variables

Detection of recombinant chIL-7 in the culture medium using Western blot
Detection of intracellular chIL-7 in the transfected cells using Western blot

control variables

Lysis buffer (5 mM Tris-HCl, 25 mM KCl, 2 mM EGTA, 2 mM EDTA, 1% NP-40, 150 mM NaCl and protease inhibitors)

controls

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

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