This is a knockout-validated antibody summary, based on the publication "Intracellular mechanisms of molecular recognition and sorting for transport of large extracellular matrix molecules", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG2bλ

Company: Enzo Life Sciences

Antibody: HSP47

Catalog number: ADI-SPA-470

Summary: Mouse monoclonal IgG2bλ against native rat HSP47. Reacts with human, mouse, rat, bovine, chicken, dog, guinea pig, hamster, monkey, porcine, rabbit, and sheep. Suitable for western blot and immunohistochemistry.

Western blot | Immunocytochemistry

Hsp47+/+ and Hsp47−/− MEFs. Cell lysates were extracted using M-PER (Thermo Fisher Scientific) containing protease inhibitors at 4 °C.

WB: PBS solution containing 5% (vol/vol) skim milk.

IC: 1:50 dilution.

WB: Alkaline phosphatase-conjugated anti-mouse and (A5781; Sigma-Aldrich).

IC: 1:1,000 dilution Alexa Fluor 568-conjugated anti-mouse IgG (A-11019; Thermo Fisher Scientific) in the dark for 30 min.

WB: 5-bromo-4-chloro-3- indolyl phosphate and Nitro blue tetrazolium.

IC: Zeiss AxioVert 200 microscope with ApoTome, and micrographs were recorded digitally using the AxioVision software (version 4.6).

Hsp47+/+ and Hsp47−/− MEFs were stained against anti-Hsp47. Total soluble proteins from equal cell numbers in Hsp47+/+ and Hsp47−/− MEFs were extracted and blotted by using anti-GAPDH and anti-Hsp47. Please see Figure 5a and 6c in the article [1].

The same clone (M16.10A1) is sold as Novus Biologicals NBP1-97491; LifeSpan Biosciences LS-B3677; Enzo Life Sciences ADI-SPA-470-D, ADI-SPA-470-F, ADI-SPA-470-J.