Laminin β-2 Antibody (H-1): sc-133241

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:200) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:200) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Five known LMα chains, three known LMβ chains and three known LMγ chains in HLE B-3 cell lysate, HLE B-3 cell culture supernatant and HLE B-3 BMs harvested at days 0, 3, 6, 9 and 12, and human ALC lysate were analyzed by western blotting, as described previously (28). Briefly, antigen sources inclding HLE B-3 cell lysate, culture supernatant or protein lysate of human ALCs were mixed with 2X sample buffer, boiled for 2 min and separated by SDS-PAGE. The separated proteins were transferred to a polyvinylidene difluoride membrane (EMD Millipore, Billerica, MA, USA). Following blocking with 5% skimmed milk in Tris-buffered saline containing 0.05% Tween-20 (TBS-T) for 1 h at room temperature, membranes were incubated with the aforementioned primary antibodies diluted in solution 1 (Toyobo Life Science, Osaka, Japan) at 4°C overnight. Following washes with TBS-T, membranes were incubated with HRP-conjugated goat anti-mouse IgG (1:5,000; cat no. 31437; Thermo Fisher Scientific, Inc.), HRP-conjugated rabbit anti-goat IgG (1:5,000; cat no. 31433; Thermo Fisher Scientific, Inc.) or HRP-conjugated goat anti-rabbit IgG (1:5,000; cat no. 31463; Thermo Fisher Scientific, Inc.) diluted in solution 2 (Toyobo Life Science) for 1 h at room temperature. Under identical experimental conditions, normal rabbit IgG, normal mouse IgG or normal goat IgG (all 1:200; cat nos. sc-2027, sc-2025 and sc-2028; Santa Cruz Biotechnology, Inc.) were used as isotype controls. Finally, the antibody-antigen complex was visualized using an Enhanced Chemiluminescent kit (Beyotime Institute of Biotechnology).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Western blotting Laminin subunit Beta-2 using Laminin β-2 Antibody (H-1): sc-133241 from Santa Cruz Biotechnology.

Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for Laminin β-2 Antibody (H-1): sc-133241 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting Laminin subunit Beta-2 using Laminin β-2 Antibody (H-1): sc-133241 from Santa Cruz Biotechnology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.