Stat3 antibody | knockout validation | Cell Signaling 9139

This is a knockout-validated antibody summary, based on the publication "Loss of signal transducer and activator of transcription 3 impaired the osteogenesis of mesenchymal progenitor cells in vivo and in vitro", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Stat3 antibody | knockout validation | Cell Signaling 9139 figure 1
Figure 1. Proteins were extracted from P0 forelimbs. Western blot analysis revealed significant decrease of Stat3 protein expression in Stat3 CKO compared with control. From [1].
Antibody information

Mouse monoclonal IgG2a

Company: Cell Signaling

Antibody: Stat3

Catalog number: 9139

Summary: Mouse monoclonal antibody against a synthetic peptide centered around amino acid Gln692 of human Stat3. Reacts with human, mouse, rat, and monkey. Suitable for western blot, immunoprecipitation (IP), immunohistochemistry (paraffin), immunofluorescence (immunocytochemistry), flow cytometry and Chromatin IP.

Stat3 antibody | knockout validation | Cell Signaling 9139 figure 2
Figure 2. IHF staining showed the loss of Stat3 expression in bone tissues on Stat3 CKO (ulna section of P0 mice). From [1].
Validation Method

Western blot | Immunohistochemistry

Sample

Control and Stat3 CKO mouse hindlimbs and bone tissues.

Blocking agent

WB: 5% BSA for 1 h.

IHC: normal goat serum (NGS) for 1 h.

Primary incubation

WB: 1:1,000 dilution at 4 °C overnight.

IHC: 1:500 dilution overnight at 4 °C.

Secondary incubation

WB: 1:4,000 dilution anti-mouse IgG HRP-linked antibody (CST, 7076) for 1 h at room temperature.

IHC: 1:500 Alexa Fluor-594 conjugated secondary antibody (Beyotime, Shanghai, China) for 1 h at room temperature.

Detection

WB: Chemiluminescence kit (FudeBio, China).

IHC: Inversion fluorescence microscope (Zeiss, Oberkochen, Germany).

References
  1. Huang Z, Feng J, Feng X, Chan L, Lu J, Lei L, et al. Loss of signal transducer and activator of transcription 3 impaired the osteogenesis of mesenchymal progenitor cells in vivo and in vitro. Cell Biosci. 2021;11:172 pubmed publisher