JNK2 antibody | knockout validation | Cell Signaling 9258

This is a knockout-validated antibody summary, based on the publication "HSF1 critically attunes proteotoxic stress sensing by mTORC1 to combat stress and promote growth", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: JNK2

Catalog number: 9258

Summary: Rabbit monoclonal IgG against human JNK2/MBP fusion protein. Reacts with human, mouse, rat, hamster, monkey, and mink. Suitable for western blot.

Validation Method

Western blot

Sample

Control and Jnk2-/- primary MEFs. Whole-cell protein extracts were prepared in cold cell-lysis buffer (100 mM NaCl, 30 mM Tris-HCl pH 7.6, 1% Triton X-100, 20 mM sodium fluoride, 1 mM EDTA, 1 mM sodium orthovanadate, and Halt™ protease inhibitor cocktail from Thermo Scientific).

Primary incubation

1:1,000 dilution in wash buffer overnight at 4°C.

Secondary incubation

1: 2,500 dilution peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories) at room temperature for 1 hr

Detection

Signals were visualized by SuperSignal West chemiluminescent substrate (Thermo Scientific) followed by exposure to films.

Figure

Primary Jnk1-/- or Jnk2-/- MEFs were treated with 200 nM MG132 for 6 hr and analyzed with the indicated antibodies by western blot. JNK2 antibodies recognized both p54 and p46 isoforms. Please see Figure 2d in the article [1].

References
  1. Su K, Cao J, Tang Z, Dai S, He Y, Sampson S, et al. HSF1 critically attunes proteotoxic stress sensing by mTORC1 to combat stress and promote growth. Nat Cell Biol. 2016;18:527-39 pubmed publisher