Anti-TGF beta 1 antibody [TB21] (ab190503)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
--These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.	-Mouse (1:500) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Upstream tips
--These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Mouse (1:500) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Publication protocol

The enucleated eyes from each mouse were lysed and the total proteins were extracted as previously described (20), and the protein concentration was quantified using a bicinchoninic acid (BCA) protein assay kit (Beyotime Institute of Biotechnology, Haimen, China). Proteins (40 µg) in each sample were loaded and separated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, Bedford, MA, USA). The membranes were then blocked with 5% non-fat milk overnight and probed with anti-p35 (ab66064), anti-IL-10 (ab134742), anti-TFG-β (ab190503), anti-IFN-γ (ab24324), anti-IL-12 (ab89889) and anti-IL-17 (ab77171) primary antibodies (1:500 dilution; all from Abcam, Cambridge, MA, USA) at 4°C overnight, followed by incubation with secondary anti-rabbit IgG-HRP antibody (A0208) or anti-mouse IgG-HRP antibody (A0216) (1:5000 dilution; Beyotime Institute of Biotechnology) at 37°C for 45 min after washing twice with TBST. The target bands were visualized using enhanced chemiluminescence (ECL) solution (Qihai Biotec, Shanghai, China) and analyzed using Gel-Pro-Analyzer software (Media Cybernetics, Bethesda, MD, USA). β-actin served as an internal control.

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Manufacturer protocol

Download the product protocol from Abcam for Anti-TGF beta 1 antibody [TB21] (ab190503) below.

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