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Human Haptoglobin ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Haptoglobin in Serum, Heparin Plasma, Citrate plasma, EDTA Plasma, Urine, Saliva, Cell culture supernatant samples.

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Images

Sandwich ELISA - Human Haptoglobin ELISA Kit (AB309180), expandable thumbnail
  • Sandwich ELISA - Human Haptoglobin ELISA Kit (AB309180), expandable thumbnail
  • Sandwich ELISA - Human Haptoglobin ELISA Kit (AB309180), expandable thumbnail
  • Sandwich ELISA - Human Haptoglobin ELISA Kit (AB309180), expandable thumbnail

Key facts

Detection method

Colorimetric

Sample types

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

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

156 - 10000 ng/mL

Assay time

1h 30m

Sensitivity

= 51.9 ng/mL

Reactivity data

Application

sELISA

Reactivity

Reacts

Dilution info

-

Notes

-

Target data

Function

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.The uncleaved form of allele alpha-2 (2-2), known as zonulin, plays a role in intestinal permeability, allowing intercellular tight junction disassembly, and controlling the equilibrium between tolerance and immunity to non-self antigens.

Alternative names

What's included?

1 x 96 Tests
Components
10X Human Haptoglobin Capture Antibody
1 x 600 µL
10X Human Haptoglobin Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
Antibody Diluent 4BI
1 x 6 mL
Human Haptoglobin Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 50 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human Haptoglobin ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Haptoglobin in Serum, Heparin Plasma, Citrate plasma, EDTA Plasma, Urine, Saliva, Cell culture supernatant samples.

Key facts

Detection method

Colorimetric

Sample types

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

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

156 - 10000 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Sensitivity

= 51.9 ng/mL

General recovery

107%

Precision

Intra assay

Sample

Serum

n

5

mean

-

SD

-

C.V.

1.8

Inter assay

Sample

Serum

n

3

mean

-

SD

-

C.V.

4.4

Recovery

Sample specific recovery

Sample type

Serum

Average %

= 107

Range

104 - 113 %

Sample type

Heparin Plasma

Average %

= 105

Range

99 - 111 %

Sample type

Citrate plasma

Average %

= 104

Range

99 - 110 %

Sample type

EDTA Plasma

Average %

= 107

Range

102 - 120 %

Sample type

Urine

Average %

= 102

Range

101 - 105 %

Sample type

Saliva

Average %

= 105

Range

101 - 113 %

Sample type

Cell culture supernatant

Average %

= 104

Range

97 - 113 %

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Human Haptoglobin SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Haptoglobin protein in human serum, plasma, saliva and cell culture supernatant. Quantitate Human Haptoglobin with 51.9 ng/mL sensitivity.

SimpleStep ELISA® technology 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. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

Supplementary info

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

Activity summary

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.

Associated diseases and disorders

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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4 product images

  • Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180), expandable thumbnail

    Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)

    Serum of ten individual healthy human male donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Haptoglobin concentration was determined to be 3.25 mg/mL with a range of 0.075 – 12.65 mg/mL. 

  • Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180), expandable thumbnail

    Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)

    Example of human Haptoglobin standard curve. Background-subtracted data values (mean +/- SD) are graphed. 

  • Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180), expandable thumbnail

    Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)

    Interpolated concentration of native Haptoglobin was measured in duplicate at different sample concentrations. Undiluted samples are as follows: urine 25%, saliva 50%, and HepG2 cell culture supernatant 12.5%.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS. 

  • Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180), expandable thumbnail

    Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)

    Interpolated concentration of native Haptoglobin was measured in duplicate at different sample concentrations. Undiluted samples are as follows: serum 1:2,000, plasma – Citrate 1:1,000, plasma – EDTA 1:1,000, and plasma – Heparin 1:2,000.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS. 

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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