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APOPT1 Antibody - C-terminal region (ARP69014_P050)

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ARP69014_P050-FITC Conjugated

ARP69014_P050-HRP Conjugated

ARP69014_P050-Biotin Conjugated

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128/05/2018 19:29
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HEK-293 Cell lysates in WB

Submitted by: Anonymous

APOPT1

1.             Sample Type/Lane Description:

Human HEK293 Cell Lysates

Lane 1: 40 ug untreated HEK293 cells (control)

Lane 2: 40 ug HEK293 cells + OA (Oligomycin + AntimycinA)

Lane 3: 40 ug HEK293 cells + CCCP

Lane 4: 40 ug HEK293 cells + Staurosporine

2.      Primary antibody dilution:

1:500

3.    Secondary antibody and dilution:

HRP-Goat anti-Rabbit IgG - 1:10,000

4.     Protocol:

HEK293 control cells, as well as cells treated with OA, CCCP, Staurosporine were lysed using a Triton X-100 based lysis buffer (1% Triton X-100, 10% glycerol, 150 mM NaCl, 20 mM Tris (pH 7.5), 2 mM EDTA) in the presence of a protease inhibitor mix. 40 μg of whole cell extract was mixed with 2 × SDS-sample buffer and boiled for 5 min. The samples were resolved on 12% SDS-PAGE and electro-transferred PVDF membranes using a semi-dry cell transfer blot. 4% nonfat dry milk in TBST buffer (25 mM Tris–HCl, pH 8.0, 125 mM NaCl, 0.1% Tween 20) was used to block nonspecific binding of the membrane. The membrane was incubated overnight at 4oC with TECPR1 (1:500 in TBST+ 4%Milk) primary antibodies followed by the specific secondary peroxidase-conjugated antibodies. The membrane was visualized by enhanced chemiluminescence (ECL) and developed using X-ray film.

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