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AB272472

Mouse Haptoglobin ELISA Kit

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Mouse Haptoglobin ELISA Kit is a single-wash 90-min Simplestep used to quantify Mouse Haptoglobin with a sensitivity of 3.91 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

Haptoglobin, Hp

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ELISA - Mouse Haptoglobin ELISA Kit (AB272472)
  • ELISA

Supplier Data

ELISA - Mouse Haptoglobin ELISA Kit (AB272472)
Sandwich ELISA - Mouse Haptoglobin ELISA Kit (AB272472)
  • sELISA

Supplier Data

Sandwich ELISA - Mouse Haptoglobin ELISA Kit (AB272472)

Interpolated concentrations of native Haptoglobin in mouse serum and plasma samples.

The concentrations of Haptoglobin were measured in duplicates, interpolated from the Haptoglobin standard curves and corrected for sample dilution. Undiluted samples are as follows : serum 1 : 250,000, plasma (citrate) 1 : 1,000,000, plasma (EDTA) 1 : 250,000, and plasma (heparin) 1 : 250,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Haptoglobin concentration was determined to be 271.1 μg/mL in serum, 836.8 μg/mL in plasma (citrate), 265.2 μg/mL in plasma (EDTA), and 282.0 μg/mL in plasma (heparin).

Sandwich ELISA - Mouse Haptoglobin ELISA Kit (AB272472)
  • sELISA

Supplier Data

Sandwich ELISA - Mouse Haptoglobin ELISA Kit (AB272472)

Example of mouse Haptoglobin standard curve in Sample Diluent NS.

The Haptoglobin standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Mouse Haptoglobin ELISA Kit (AB272472)
  • sELISA

Supplier Data

Sandwich ELISA - Mouse Haptoglobin ELISA Kit (AB272472)

Interpolated concentrations of native Haptoglobin in mouse tissue culture supernatant and urine samples.

The concentrations of Haptoglobin were measured in duplicates, interpolated from the Haptoglobin standard curves and corrected for sample dilution. Undiluted samples are as follows : liver supernatant 1 : 200 and urine 1 : 160. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Haptoglobin concentration was determined to be 211.7 ng/mL in liver supernatant and 109.3 ng/mL in urine.

Key facts

Detection method

Colorimetric

Sample types

Urine, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Reacts with

Mouse

Assay type

Sandwich (quantitative)

Sensitivity

= 3.91 pg/mL

Range

18.75 - 1200 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Mouse Haptoglobin ELISA Kit ab272472 is a rapid single-wash 90-min Sandwich ELISA to measure Mouse Haptoglobin in cell culture supernatant, citrate plasma, EDTA plasma, heparin plasma, serum, urine. This SimpleStep sensitivity is 3.91 pg/mL.

How the assay works

Mouse Haptoglobin SimpleStep ELISA®employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Mouse Haptoglobin ELISA Kit ab272472 protocol summary

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-Haptoglobin antibody [EPR22856-71] - BSA and Azide free (Capture) ab272756
- Anti-Haptoglobin antibody [EPR22856-277] - BSA and Azide free (Detector) ab272756

Haptoglobin, encoded by the HP gene and produced mostly by hepatic cells, is a major acute phase protein. Haptoglobin is known to bind to free hemoglobin released from erythrocytes with high affinity and inhibits its oxidative activity to allow hepatic recycling of the heme iron. Biosynthesis of haptoglobin can also occur in adipose and lung tissues to provide antioxidant and antimicrobial activity. Studies have shown haptoglobin to play a role in stimulation of angiogenesis and changes in the measured concentrations in serum may associate with certain inflammations, infections, and hemolytic conditions. Human, rat, cow, monkey share 81%, 90%, 76%, and 80% sequence homology with mouse haptoglobin, respectively.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.8" }, { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.5" } ]
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Recovery

[ { "sample": "Cell culture supernatant", "range": "113 - 117 %", "average": "= 115" }, { "sample": "Serum", "range": "107 - 113 %", "average": "= 110" }, { "sample": "EDTA Plasma", "range": "103 - 108 %", "average": "= 106" }, { "sample": "Urine", "range": "114 - 118 %", "average": "= 116" }, { "sample": "Heparin Plasma", "range": "104 - 113 %", "average": "= 109" }, { "sample": "Citrate plasma", "range": "110 - 111 %", "average": "= 111" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Haptoglobin (Hp) also known as Hp protein is a glycoprotein with a molecular weight ranging from 85 to 100 kDa varying due to its ability to form different dimeric and multimeric structures. It is primarily synthesized in the liver and circulating in the plasma. Mechanically haptoglobin binds free hemoglobin released from erythrocytes preventing oxidative damage and hemoglobin-driven deleterious effects. Haptoglobin's binding to hemoglobin forms a haptoglobin-hemoglobin complex that gets cleared by the CD163 receptor on macrophages mainly in the liver and spleen.
Biological function summary

Haptoglobin functions as an antioxidant and an acute phase protein playing roles in inflammation and immune response. It is an essential component of the antioxidant defense system reducing oxidative stress by binding free hemoglobin. Additionally haptoglobin can participate in complex formation with hemoglobin to facilitate the clearance of excess free hemoglobin from circulation. Its expression increases during inflammation under the regulation of cytokines like IL-6.

Pathways

The haptoglobin protein is part of the hemoglobin clearance pathway integral for iron metabolism and homeostasis. This pathway ensures the safe removal of hemoglobin from the blood helping to prevent kidney damage. Haptoglobin is related to other proteins such as CD163 and the liver-derived hepatocellular proteins involved in the clearance process. Also haptoglobin is linked to antioxidant pathways collaborating with oxidative stress regulators to manage reactive oxygen species levels.

Changes in haptoglobin levels are associated with hemolytic anemia and inflammatory conditions. Low haptoglobin levels often suggest hemolytic anemia due to its rapid binding with free hemoglobin in the bloodstream. During inflammation and infections 'anti-Hp' tests (haptoglobin tests) may show increased haptoglobin levels as a response to cytokine signaling. Moreover haptoglobin polymorphisms have a connection with ischemic stroke susceptibility where its isoforms show differing efficiencies in binding free hemoglobin. These findings highlight haptoglobin's relevance as a biomarker for these conditions and illustrate its interaction with hematologic and immune proteins in disease contexts.
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Product protocols

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Target data

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 antioxidant, has antibacterial activity and plays a role in modulating many aspects of the acute phase response. Hemoglobin/haptoglobin complexes are rapidly cleared by the macrophage CD163 scavenger receptor expressed on the surface of liver Kupfer cells through an endocytic lysosomal degradation pathway (By similarity).
See full target information Hp
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websiteProtocolBooklet
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