This is a knockout-validated antibody summary, based on the publication "Hematopoietic Stem and Progenitor Cell Proliferation and Differentiation Requires the Trithorax Protein Ash2l", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Serial tissue sections from fixed sterna stained with anti-Ash2l or anti-H3K4me3 antibodies as indicated. Western blot analysis of proteins of BM cell lysates (from control and Ash2l KO mice). Upper and lower halves of the blot were probed for Ash2l and actin, respectively. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: ASH2L

Catalog number: 5019S

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to the sequence of human ASH2L protein. Reacts with human, mouse, rat, and monkey. It is predicted to react based on 100% sequence homology with D. melanogaster. Suitable for western blot, immunoprecipitation, and immunofluorescence/immunocytochemistry.

Western blot | Immunohistochemistry

Sterna and BM cells from control and Ash2l KO mice. WB: Cells were lysed on ice in RIPA buffer (10 mM Tris/HCl, pH 7.4, 150 mM NaCl, 1% Nonidet P-40, 1% deoxycholic acid, 0.1% SDS) containing a protease inhibitor cocktail and 10 mM sodium butyrate. IHC: sternum bones were fixed in 4% formaldehyde, decalcified in 10% EDTA (pH 7.4), embedded in paraffin and cut into 2 µm sections.

IHC: following peroxidase-blocking solution, the sections were treated with 2.5% normal horse serum for 30 min and incubated at room temperature for 30 min.

WB: 1:2,000 dilution in 5% low-fat milk, 0,05% Tween20 in PBS. IHC: 1:750 dilution in antibody diluent.

WB: 1:2,000 dilution peroxidase anti-rabbit antibodies. IHC: ImmPress peroxidase kit was used (20 min), followed by incubation with DAB (10 min).

WB: SuperSignal Femto Substrate. IHC: NanoZoomer from Hamamatsu Photonics and visualized using the NDP.view2 software.