Detection of Human alpha 2‑Macroglobulin by Western Blot.
Western blot shows human serum. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human alpha 2‑Macroglobulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1938) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for alpha 2‑Macroglobulin at approximately 180 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
alpha 2‑Macroglobulin was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human alpha 2‑Macroglobulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1938) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human alpha 2‑Macroglobulin by Simple WesternTM.
Simple Western lane view shows human serum, loaded at 0.2 mg/mL. A specific band was detected for alpha 2‑Macroglobulin at approximately 178 kDa (as indicated) using 1 µg/mL of Goat Anti-Human alpha 2‑Macroglobulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1938) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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 2-Macroglobulin
Human alpha 2-macroglobulin (h alpha 2M) is a serum glycoprotein that has sequence similarity to other members of the alpha 2M family including complement components C3, C4 and C5 (1). alpha 2M is synthesized as a polypeptide of 1474 amino acids with a signal peptide (23 residues) (2). The mature protein is a tetramer (720 kDa) of 4 identical subunits (180 kDa), which form two disulfide bond-linked dimers. As a general and irreversible protease inhibitor implicated in many processes, alpha 2M is able to inhibit all four classes of proteases by a unique trapping mechanism. The bait region of h alpha 2M (residues 690‑728) contains specific cleavage sites for different proteases. The cleavage of the bait region by a protease induces a conformation change in alpha 2M, which then traps and forms a covalent bond with the protease. The trapped protease remains active against small peptide substrates but loses its ability to interact with large protein substrates or inhibitors.
Sottrup-Jensen, L. et al. (1985) Proc. Natl. Acad. Sci. USA 82:9.
Kan, C.C. et al. (1985) Proc. Natl. Acad. Sci. USA 82:2282.
Entrez Gene IDs:
A2M; alpha 2Macroglobulin; alpha 2-Macroglobulin; alpha-2-M; alpha-2-macroglobulin; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5; CPAMD5; CPAMD5DKFZp779B086; FWP007; S863-7
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