Human Haptoglobin EZ-Set™ ELISA Kit (DIY Antibody Pairs)

Haptoglobin (HP), is a protein that in humans is encoded by the HP gene. Haptoglobin, a plasma glycoprotein that binds free hemoglobin, has a tetrameric structure of 2 alpha and 2 beta polypeptides that are covalently associated by disulfide bonds. Haptoglobin is homologous to serine proteases of the chymotrypsinogen family. In the chimpanzee, there are 3 genes in the haptoglobin family (haptoglobin, HP; haptoglobin-related, HPR; and haptoglobin-primate, HPP), whereas only 2 genes exist in humans (HP and HPR) . There are similarities between the primary structures of the alpha chain and of light chains of gamma globulins. The alpha haptoglobin locus is on the long arm of chromosome 16, where its exact cytogenetic location is 16q22.2. A major function of haptoglobin is to bind hemoglobin (Hb) to form a stable Hp-Hb complex and thereby prevent Hb-induced oxidative tissue damage. Haptoglobin is an unusual secretory protein in that it is proteolytically processed in the endoplasmic reticulum and not in the Golgi. The human haptoglobin HP*2 allele contains a 1.7-kb intragenic duplication that arose after a unique nonhomologous recombination between the prototype HP*1 alleles.

As a result of hemolysis, hemoglobin is found to accumulate in the kidney and is secreted in the urine. Haptoglobin captures, and combines with free plasma hemoglobin to allow hepatic recycling of heme iron and to prevent kidney damage. Haptoglobin also acts as an Antimicrobial; Antioxidant, has antibacterial activity and plays a role in modulating many aspects of the acute phase response. Hemoglobin/haptoglobin complexes are rapidely cleared by the macrophage CD163 scavenger receptor expressed on the surface of liver Kupfer cells through an endocytic lysosomal degradation pathway.

Catalog number Description Quantity Storage of opened/reconstituted material EZ1702-CA Mouse anti-human Haptoglobin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1702-DA Biotinylated goat anti-human Haptoglobin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1702-ST Lyophilized recombinant human Haptoglobin standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1702-CA Mouse anti-human Haptoglobin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1702-DA Biotinylated goat anti-human Haptoglobin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1702-ST Lyophilized recombinant human Haptoglobin standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1702-CA Mouse anti-human Haptoglobin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1702-CA Mouse anti-human Haptoglobin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1702-DA Biotinylated goat anti-human Haptoglobin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) EZ1702-DA Biotinylated goat anti-human Haptoglobin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1702-ST Lyophilized recombinant human Haptoglobin standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EZ1702-ST Lyophilized recombinant human Haptoglobin standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 O.D. 0.070 0.332 0.526 0.998 1.329 1.720 1.961 2.389 Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 O.D. 0.070 0.332 0.526 0.998 1.329 1.720 1.961 2.389 Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 O.D. 0.070 0.332 0.526 0.998 1.329 1.720 1.961 2.389 O.D. 0.070 0.332 0.526 0.998 1.329 1.720 1.961 2.389

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)