Shh antibody | knockout validation | DSHB 5E1

This is a knockout-validated antibody summary, based on the publication "Epithelial heparan sulfate regulates Sonic Hedgehog signaling in lung development", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Mouse monoclonal IgG1

Company: DSHB

Antibody: Shh

Catalog number: 5E1

Summary: Mouse monoclonal IgG1 against a synthetic peptide corresponding to rat sonic hedgehog N-terminal SSH fragment a.a. 1-198. Reacts with chicken, human, mouse, quail, rabbit, and rat. Suitable for western blot, immunofluorescence, immunohistochemistry, flow cytometry and function blocking experiments.

Validation Method

Western blot

Sample

Control Shhcre/+ lungs and Shh null lungs (Shhcre/cre). Embryos were fixed in 4% paraformaldehyde at 4°C overnight and embedded in paraffin for sectioning.

Primary incubation

1:20 dilution.

Secondary incubation

ABC immunoperoxidase detection systems (Vector PK-6200 Laboratories) was used to introduce HRP. The Cyanine 3 Tyramide Signal detection was performed according to the guidelines of TSA amplification system (PerkinElmer).

Detection

Zeiss LSM 780 laser-scanning confocal microscope under the same laser exposure between control and mutant lungs.

Figure

Immunofluorescent staining of Shhcre/+ lungs and Shh null lungs (Shhcre/cre) using 5E1 antibody. The cluster signal were found both in the epithelium and the mesenchyme of the control lungs with strong stainings in the basement membrane. Shh null lungs were negative for the signal. Scale bar: A and B, 20μm. From [1].

References
  1. He H, Huang M, Sun S, Wu Y, Lin X. Epithelial heparan sulfate regulates Sonic Hedgehog signaling in lung development. PLoS Genet. 2017;13:e1006992 pubmed publisher