Detection of Mouse and Rat Integrin beta 5 by Western Blot.
Western blot shows lysates of CMT-93 mouse rectal carcinoma cell line and C6 rat glioma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Mouse Integrin beta 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8035) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Integrin beta 5 at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Integrin beta 5 in bEnd.3 Mouse Cell Line by Flow Cytometry.
bEnd.3 mouse endothelioma cell line was stained with Sheep Anti-Mouse/Rat Integrin beta 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8035, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127).
Integrin beta 5 in Mouse Brain.
Integrin beta 5 was detected in perfusion fixed frozen sections of mouse brain using Sheep Anti-Mouse/Rat Integrin beta 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8035) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010). Specific staining was localized to glial cells. View our protocol for Fluorescent IHC Staining of Frozen 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.
Background: Integrin beta 5
The beta 5 integrin is a 95-110 kDa member of the integrin beta chain family, integrin superfamily of molecules. It is expressed by select cell types, including microglia, keratinocytes, fibroblasts, endothelial cells, adhesive monocytes and macrophages, retinal pigment epithelium, immature dendritic cells and colonic epithelium. The Integrin beta 5 subunit forms a noncovalent heterodimer with the alpha v integrin subunit, and together they bind vitronectin, and the Thy-1 antigen. alpha v beta 5 is reported to participate in phagocytosis, and act in concert with CD81, CD36 and stabilin-2 on the cell surface. When alpha v beta 5 binds Thy-1, the heterodimer apparently interfers with cell membrane activation of TGF-beta. Intracellularly, it appears to regulate actin skeleton remodeling via focal adhesion complex formation, and may contribute to cryptic pathways associated with antigen presentation. Mature mouse Integrin beta 5 is a 775 amino acid (aa) type I transmembrane glycoprotein that contains a 696 aa extracellular domain (ECD) (aa 24-719) and a 56 aa cytoplasmic region. The ECD contains a 243 aa vWF-A domain (aa 136-378) and a Cys-rich tandem repeat region (aa 465-630). There is one splice variant that shows a 57 aa substitution for aa 760-798. This splice form does not appear to be expressed by the human Integrin beta 5 gene. Over aa 24-719, mouse Integrin beta 5 shares 91% and 97% aa sequence identity with human and rat Integrin beta 5, respectively.
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