Detects rat SIRP-alpha /CD172a in direct ELISA and Western Blot.
Monoclonal Mouse IgG2B Clone # 772734
Protein A or G purified from hybridoma culture supernatant
Mouse myeloma cell line NS0-derived recombinant rat SIRP-alpha /CD172a Lys32-Asn373 Accession # P97710
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
2.5 µg/106 cells
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Measured by its ability to neutralize SIRP alpha /CD172a-induced adhesion in mouse red blood cells. The Neutralization Dose (ND50) is typically 0.04-0.16 μg/mL in the presence of 6 μg/mL Recombinant Rat SIRP alpha /CD172a.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Rat SIRP alpha /CD172a by Western Blot. Western blot shows lysates of NR8383 rat alveolar macrophage cell line and rat brain tissue. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Rat SIRP alpha /CD172a Monoclonal Antibody (Catalog # MAB7307) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for SIRP alpha /CD172a at approximately 75-140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Adhesion Induced by SIRP alpha / CD172a and Neutralization by Rat SIRP alpha /CD172a Antibody. Recombinant Rat SIRP alpha /CD172a (Catalog # 7307-SA) supports the adhesion in mouse red blood cells in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Rat SIRP alpha /CD172a (6 μg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Rat SIRP alpha /CD172a Monoclonal Antibody (Catalog # MAB7307). The ND50 is typically 0.04-0.16 μg/mL.
Detection of SIRP alpha /CD172a in PC‑12 Rat Cell Line by Flow Cytometry. PC‑12 rat adrenal pheochromocytoma cell line was stained with Mouse Anti-Rat SIRP alpha /CD172a Monoclonal Antibody (Catalog # MAB7307, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B).
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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: SIRP alpha/CD172a
SIRP alpha (Signal regulatory protein alpha; also CD172a, Shps1 and Bit) is a variably glycosylated, 85-120 kDa member of the SIRP ‘family' of proteins. It is expressed on neurons, macrophages, monocytes, granulocytes and dendritic cells. SIRP alpha is phosphorylated/activated in response to cell adhesion. This may, or may not, involve binding to known ligands CD47, SP-A and SP-D. Following phosphorylation, SIRP alpha binds to SHP-1 and -2, resulting in the negative regulation of immune system activity. Mature rat SIRP alpha is a 478 amino acid (aa) type I transmembrane glycoprotein. It contains a 342 aa extracellular region (aa 32-373) that possesses one V-type and two C1-type Ig-like domains. Its cytoplasmic domain possesses two ITIMs that interact with protein phosphatases. There is one potential splice variant that shows a four aa insertion after Gln424. Over aa 32-373, rat SIRP alpha shares 63% and 73% aa sequence identity with human and mouse SIRP alpha, respectively.
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