Pumilio 1 antibody | knockout validation | Abcam ab92545

This is a knockout-validated antibody summary, based on two publications "Post-transcriptional regulation of mouse neurogenesis by Pumilio proteins" [1] (western blot, see figure 2c in the article), and "An Important Role of Pumilio 1 in Regulating the Development of the Mammalian Female Germline", as cited below [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Pumilio 1 antibody | knockout validation | Abcam ab92545 figure 1
Figure 1. Immunoblot analysis showing PUM1 and PUM2 protein levels in wild-type mouse ovaries (+/+) and Pum1−/− ovaries (−/−) from 6- to 10-wk-old female mice. Glyceraldehyde phosphate dehydrogenase (GAPDH) used as loading control. From [2].
Antibody information

Rabbit monoclonal IgG

Company: Abcam

Antibody: Pumilio 1

Catalog number: ab92545

Summary: Rabbit monoclonal IgG against a synthetic peptide within human Pumilio 1( aa 150-250). Reacts with mouse, rat, and human. Suitable for western blot, immunocytochemistry /immunofluorescence, immunohistochemistry (paraffin) and flow cytometry. Unsuitable for immunoprecipitation.

Validation Method

Western blot

Sample

Wild-type and Pum1−/− mouse ovaries. Samples were prepared in M-Per Mammalian protein extraction reagent (Thermo Scientific) according to manufacturer’s instructions.

Blocking agent

5% milk TBST solution for 30 min.

Primary incubation

1:1,000 dilution overnight at 4 C.

Secondary incubation

1:4,000 dilution anti-rabbit conjugated horseradish peroxidase (Life Technologies) in 5% milk TBST for 1 h at room temperature.

Detection

Chemiluminescent signals were detected using Supersignal West Dura extended substrate (Thermo Scientific) and radiographic film.

References
  1. Zhang M, Chen D, Xia J, Han W, Cui X, Neuenkirchen N, et al. Post-transcriptional regulation of mouse neurogenesis by Pumilio proteins. Genes Dev. 2017;31:1354-1369 pubmed publisher
  2. Mak W, Fang C, Holden T, Dratver M, Lin H. An Important Role of Pumilio 1 in Regulating the Development of the Mammalian Female Germline. Biol Reprod. 2016;94:134 pubmed publisher