Vimentin polyclonal antibody

Total cellular proteins were extracted by RIPA lysis buffer (NCM Biotech, Suzhou, China) combined with protease inhibitor and phosphatase inhibitor (NCM Biotech, Suzhou, China). The BCA protein detection kit (Beijing Biotech Co., Ltd., China) was used to determine the protein concentration. We added native gel sample loading buffer (NCM Biotech, Suzhou, China) to the protein samples and heated them to denature them. The proteins were separated in gel prepared by ExpressCast PAGE Preparation Kit and then transferred onto 0.45 μm polyvinylidene fluoride (PVDF) membranes (Millipore, Darmstadt, Germany). Then, the membrane was blocked at room temperature for 2 h and incubated with the primary antibody at 4 °C overnight. Then, we used the second antibody (Abmart, Shanghai, China) to incubate the membrane at room temperature for 1 h. Enhanced Chemiluminescent (NCM Biotech, Suzhou, China) was used to show the strip. The primary antibodies we used included: FAM50A Rabbit mAb (ABclonal Technology, Wuhan, China), E-cadherin Polyclonal antibody (Proteintech, Chicago, IL, USA), N-cadherin Rabbit mAb (CST, Boston, MA, USA), Vimentin Polyclonal Antibody (Proteintech, Chicago, IL, USA), β-Tubulin mouse mAb (Abmart, Shanghai, China) and GAPDH antibody (NCM Biotech, Suzhou, China).

Mouse flow cytometry antibodies CD16/CD32 (553141), CD45 APC-Cy7 (557659), CD11b PE (553311), Ly-6G and Ly-6C FITC (553127), and flow cytometry buffer (Stain Buffer FBS) were purchased from BD Pharmingen (United States). The red blood cell lysate was purchased from Biyuntian Company (China). Type II lung epithelial cell isolation and identification antibody Biotinylated anti-mouse CD45, Biotinylated anti-mouse TER119, Anti-mouse EpCAM-APC were purchased from eBioscience (United States). Biotinylated anti-mouse Integrin β4, Biotinylated anti-mouse CD31 were purchased from Biolegend (United States). Streptavidin-PE, Biotinylated anti-mouse CD16/32 were purchased from BD Pharmingen (United States). Dynabeads^® MyOne^TM streptavidin T1 magnetic beads, Rabbit anti-pro-SP-C, and DAPI were purchased from Thermo Fisher Scientific (United States). Dispase II, low melting point agarose gel, DNase I were purchased from Sigma (United States). VWF, VWFpp polyclonal antibody and vimentin polyclonal antibody were purchased from Proteintech (United States). Mcp1 Monoclonal antibody was purchased from Proteintech (United States). Mouse antibodies against CD63, Tsg101, and CD81 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States). Rabbit antibodies against PTEN, Phospho-NF-κB p65 Phospho-Akt, Calnexin were purchased from Cell Signaling Technology (MA, United States).

Antibodies were listed as follows: Phospho‐Rb (Ser807/811) (D20B12) XP Rabbit mAb (Cell Signaling Technology, Cat: 8516), Cyclin D1 (E3P5S) XP Rabbit mAb (Cell Signaling Technology, Cat: 55506), GAPDH (D16H11) XP Rabbit mAb (Cell Signaling Technology, Cat: 5174), Vimentin Polyclonal Antibody (Proteintech, Cat: 10366‐1‐AP), N‐Cadherin Polyclonal Antibody (Proteintech, Cat: 22018‐1‐AP), Anti‐non‐muscle Myosin IIA antibody [EPR22933‐9] (Abcam, Cat: ab238131), Anti‐MYH9 Polyclonal Antibody (Beijing Solarbio Science & Technology Co., Ltd.), Anti‐Hsp90 antibody [EPR16621‐67] (Abcam, Cat: ab203126), Anti‐E Cadherin antibody [SP64] (Abcam, Cat: ab227639).