Purified anti-mouse/rat/human FOXP3 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Mouse (1:500)

Protocol tips
Mouse (1:500)

Publication protocol

Single cells from injured muscles were obtained by enzymatic digestion of the tissue with collagenase type IV (Sigma Aldrich) (0.5 mg/ml) and dispase (Invitrogen) (3.5 mg/ml) at 37°C for 40 min. CD3+ cells were then isolated by magnetic sorting (CD3ε MicroBead Kit, Milteniy) following manufacturer’s instruction. Isolated CD3+ cells were lysed in RIPA buffer and protease inhibitors cocktail (Sigma Aldrich). Lysates were cleaned by centrifugation at 16,000 x g for 5 min at 4°C. For Western blot analysis, equal amounts of protein (20 μg) were resolved by SDS polyacrylamide gel electrophoresis and transferred onto Immobilon-P (Millipore). Antigens were detected using mouse anti-FOXP3 (1:500, clone 150D, Biolegend), rat anti-RORγT (1:250, clone B2D, eBioscience), mouse anti-GATA3 (1:200, clone HG3-31, Santa Cruz biotec), mouse anti-TBET (1:200, clone 4B10, Santa Cruz biotec), mouse monoclonal anti-β-actin (1:10000, clone AC15, Sigma Aldrich). All antibodies were diluted in TBST containing 5% non-fat milk. Incubation was performed 2 hours at room temperature for primary antibodies and 1 hour at room temperature for second step reagents. Primary antibodies were revealed with HRP-conjugated secondary antibodies (1:1000, GE Healthcare Europe GmbH) and a chemiluminescence kit (ECL, western blotting detection reagents, GE Healthcare Europe GmbH). When necessary, membranes were stripped with 0.1N NaOH for 10 min at room temperature.

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 FOXP3 using Purified anti-mouse/rat/human FOXP3 Antibody from BioLegend.

Manufacturer protocol

Download the product protocol from BioLegend for Purified anti-mouse/rat/human FOXP3 Antibody below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting FOXP3 using Purified anti-mouse/rat/human FOXP3 Antibody from BioLegend. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.