Detection of Human alpha 1‑Microglobulin by Western Blot.
Western blot shows lysates of human liver tissue, Hep3B human hepatocellular carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, and Huh‑7 human hepatoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human alpha 1‑Microglobulin Monoclonal Antibody (Catalog # MAB7724) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for alpha 1‑Microglobulin at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
alpha 1-Microglobulin in HepG2 Human Cell Line.
alpha 1-Microglobulin was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Mouse Anti-Human alpha 1-Microglobulin Monoclonal Antibody (Catalog # MAB7724) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 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: alpha 1-Microglobulin
Human alpha 1-Microglobulin (alpha1-m/A1M; also protein HC) is a secreted, 31-32 kDa glycoprotein member of the lipocalin family, calycin superfamily of molecules. It is expressed by hepatocytes, keratinocytes, and endodermal derivatives in the embryo. alpha 1-Microglobulin appears to act as a heme scavenger, protecting cells and collagen against oxidative damage. It also acts as an immunosuppressant, inhibiting polyclonal lymphocyte activation and dampening granulocyte migration in response to chemokines. alpha 1-Microglobulin circulates either as a monomer, or bound to IgA, albumin or prothrombin. Human alpha 1-Microglobulin is generated through cleavage of a precursor molecule termed AMBP. This AMBP should not be confused with AMBP-1, a 120-140 kDa adrenomedullin-binding protein that is also known as Complement Factor H. The AMBP precursor contains a 19 aa signal sequence, an N-terminal 183 aa alpha 1-Microglobulin protein (aa 20-203), and a C-terminal serine protease inhibitor termed bikunin (aa 206-352). alpha 1-Microglobulin possesses one lipocalin domain (aa 42-186). Although cleavage of AMBP in the Golgi apparatus typically generates a 31 kDa alpha 1-Microglobulin and 28 kDa bikunin molecule, the 60-65 kDa AMBP precursor can also be released intact. alpha 1-Microglobulin will undergo extracellular processing, generating a 30 kDa isoform that is missing aa 199-203. There is one splice variant that shows a deletion of aa 48-57. Over aa 20-203, human alpha 1-Microglobulin shares 76% aa sequence identity with both mouse and rat alpha 1-Microglobulin.
Entrez Gene IDs:
259 (Human); 11699 (Mouse); 25377 (Rat)
A1M; alpha 1Microglobulin; alpha 1-Microglobulin; alpha-1-microglobulin/bikunin precursor; AMBP; bikunin; EDC1; HCP; HCPcomplex-forming glycoprotein heterogeneous in charge; HI30; IATIL; inter-alpha-trypsin inhibitor light chain; ITI; ITIL; ITILC; ITILgrowth-inhibiting protein 19; protein AMBP; protein HC; trypstatin; uristatin; uronic-acid-rich protein; UTI
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