Tgf beta 1 antibody

DMEM-F12 medium and foetal bovine serum (FBS) were purchased from the Gibco (Invitrogen, USA). Penicillin, streptomycin and BCA protein assay kits were purchased from the Solarbio (Beijing, China). Polystyrene plate for transwell was purchased from the Corning (New York, USA). The primary antibodies were diluted 1:2000 before use, including OLA1 (Cat. #ab229090, Abcam), GAPDH (Cat. #ab9485, Abcam), GAPDH (Cat. #ab8245, Abcam), N-cadherin (Cat. #ab18203, Abcam), E-cadherin (Cat. #ab76055, Abcam), Snail (Cat. #PA5–11923, Invitrogen), Snail + Slug (Cat. # ab180714, Abcam), Zeb-1 (Cat. # ab155249 Abcam) and SMAD2 (Cat. #5339, Cell Signaling Technology), p-SMAD2 (Cat. #18338, Cell Signaling Technology), SMAD4 (Cat. #9515P, Cell Signaling Technology). TGF-beta 1 antibody (Cat. # 21898–1-AP) was purchased from ProteinTech (Wuhan, China). All the chemical compounds were analytically pure reagents.

Protein was extracted from the rats’ ovaries using RIPA (radioimmunoprecipitation assay) lysis buffer and was quantified by using a BCA protein assay kit (Thermo Fisher, SE253117A). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was performed with 10% separation glue. Then the protein on the gel was transferred to polyvinylidene difluoride (PVDF) membrane under the condition of constant 300 mA. 5% skimmed milk dissolved in Tris-buffered saline with Tween (TBST) was used to block the PVDF membrane after transmembrane. The primary antibody was incubated overnight at 4 °C, and the secondary antibody was incubated for 1 h at room temperature. After that, an ECL kit was used to determine chemiluminescence.
The antibodies were TGF-Beta1 Antibody (Proteintech, 21898-1-AP), FSHR Antibody (Proteintech, 22665-1-AP), Anti-Smad4 (Abcam, ab40759), JNK (Phospho-Thr183/Thr183/Thr221) Rabbit pAb (ZENBIO, 384799), JNK Antibody (Proteintech, 10023-1-AP), Phospho-Smad2 (Ser467) Rabbit pAb (ZENBIO, 310079), Smad2/3 Rabbit pAb (ZENBIO, 382472), Phospho-Smad3 (Ser423/425) Rabbit pAb (ZENBIO, 382919), Smad3 Rabbit pAb (ZENBIO, 382774), TAK1 (Phospho-Ser439) Rabbit pAb(ZENBIO, 385849), TAK1 (3G1) Mouse mAb (ZENBIO, 200993), TRAF6 Rabbit pAb (ZENBIO, 385965), p38 MAPK (CST, 8690S), Phospho-p38 MAPK (CST, 4631S), GAPDH Antibody (Boster, BA2913), HRP-linked Antibody (CST, 7074).