Detection of Human Hemopexin by Western Blot.
Western blot shows lysates of human liver tissue. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Hemopexin Monoclonal Antibody (Catalog # MAB4490) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Hemopexin at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 5.
Hemopexin in Human Liver.
Hemopexin was detected in immersion fixed paraffin-embedded sections of human liver using Mouse Anti-Human Hemopexin Monoclonal Antibody (Catalog # MAB4490) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membranes of hepatocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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.
Hemopexin (HPX) is a 60 kDa plasma glycoprotein with two four-bladed beta -propeller folds. This structural motif has been found in other proteins including collagenases and provides sites for protein-protein interactions (1-3). The liver is the major synthesizing organ. Expression in the central nervous system, in the retina, and in peripheral nerves has also been observed. Hemopexin belongs to the family of the acute-phase proteins whose synthesis is induced after an inflammatory event. Hemopexin participates in maintaining and recycling the iron pool by utilizing its high binding affinity toward heme composed of protoporphyrin IX and iron. It also functions in preventing oxidation caused by heme after hemolysis. Hydrophobic heme molecules can intercalate into lipid membranes and participate in the oxidation of lipid membrane components through the Fenton reaction resulting in lipid peroxidation. Hemopexin undergoes a conformational change upon the binding of heme. The conformational change allows hemopexin to interact with a specific receptor, forming a complex which is then internalized. In the plasma, it is likely that heme binds albumin (35-55 g/L) first because of the higher concentration of albumin in plasma than hemopexin (0.5-1.2 g/L), and is then transferred to hemopexin, which has a much higher affinity (Kd ~ 1 pM) toward heme. Heme concentrations in plasma increase after hemolysis, which is associated with several pathological conditions such as reperfusion injury and ischemia.
Tolosano, E. and Altruda, F. (2002) DNA and Cell Biol. 21:297.
Mauk, M. R. et al. (2007) Nature Pro. Rep. 24:523.
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