Detection of Human ASCL2/Mash2 by Western Blot.
Western blot shows lysates of JAR human choriocarcinoma cell line and MCF‑7 human breast cancer cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human ASCL2/Mash2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6539) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ASCL2/Mash2 at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
ASCL2/Mash2 in Human Intestine.
ASCL2/Mash2 was detected in immersion fixed paraffin-embedded sections of human intestine using Sheep Anti-Human ASCL2/Mash2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6539) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hemotoxylin (blue). Lower panel shows a lack of labeling when primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. Specific staining was localized to the base of intestinal crypts. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
ASCL2 (Achaete-scute like 2; also HASH2, bHLHa45 and Mash2) is a class II member of the HLH family of transcription factors. Its predicted MW is 20 kDa. ASCL2 shows restricted expression, being limited to intestinal Lgr5+ stem cells and first trimester placental cytotrophoblasts. In the intestine, ASCL2 is under the control of Wnts and serves to maintain an epithelium stem cell pool. In the placenta, ASCL2 acts to maintain the pool of cytotrophoblasts at the expense of synctiotrophoblasts, thus promoting placental growth. Human ASCL2 is 193 amino acids (aa) in length. It contains one poly-Arg motif (aa 36-39), a DNA binding sequence (aa 53-63) and an HLH domain (aa 64-103). ASCL2 forms heterodimers with daughterless homologs (E22, E2A and HEB). Over aa 1-49, human ASCL2 shares 59% aa identity with mouse Mash2.
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