Overview

  • Product name

    Human Haptoglobin ELISA Kit
    See all Haptoglobin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 4.9%
    Inter-assay
    Sample n Mean SD CV%
    Overall 7.2%
  • Sample type

    Serum, Plasma
  • Assay type

    Competitive
  • Sensitivity

    = 0.07 µg/ml
  • Range

    0.078 µg/ml - 20 µg/ml
  • Recovery

    = 98 %

  • Assay time

    2h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human Haptoglobin in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Haptoglobin in plasma and serum.


    A Haptoglobin specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently biotinylated Haptoglobin is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Complex is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize Streptavidin-Peroxidase enzymatic reaction. TMB is catalyzed by Streptavidin-Peroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is inversely proportional to the amount of Haptoglobin captured in plate.


    The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.

  • Tested applications

    Suitable for: Competitive ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Streptavidin-Peroxidase Conjugate 1 x 80µl
    10X Diluent N Concentrate 1 x 30ml
    20X Wash Buffer Concentrate 1 x 30ml
    2X Biotinylated Human Haptoglobin (Lyophilized) 1 vial
    Chromogen Substrate 1 x 8ml
    Haptoglobin Microplate (12 x 8 well strips) 1 unit
    Haptoglobin Standard 1 vial
    Sealing Tapes 3 units
    Stop Solution 1 x 12ml
  • Research areas

  • Function

    Haptoglobin combines with free plasma hemoglobin, preventing loss of iron through the kidneys and protecting the kidneys from damage by hemoglobin, while making the hemoglobin accessible to degradative enzymes.
    Uncleaved haptoglogin, also 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.
  • Tissue specificity

    Expressed by the liver and secreted in plasma.
  • Involvement in disease

    Defects in HP are the cause of anhaptoglobinemia (AHP) [MIM:614081]. AHP is a condition characterized by the absence of the serum glycoprotein haptoglobin. Serum levels of haptoglobin vary among normal persons: levels are low in the neonatal period and in the elderly, differ by population, and can be influenced by environmental factors, such as infection. Secondary hypohaptoglobinemia can occur as a consequence of hemolysis, during which haptoglobin binds to free hemoglobin.
  • Sequence similarities

    Belongs to the peptidase S1 family.
    Contains 1 peptidase S1 domain.
    Contains 2 Sushi (CCP/SCR) domains.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • binding peptide
    • BP
    • haptoglobin alpha 1S beta
    • haptoglobin alpha 2FS beta
    • haptoglobin alpha polypeptide
    • Haptoglobin beta chain
    • haptoglobin beta polypeptide
    • HP
    • HP2ALPHA2
    • HPA1S
    • HPT_HUMAN
    • Zonulin
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab108856 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Competitive ELISA Use at an assay dependent dilution.

Images

  • Haptoglobin measured in various biological fluids showing quantity (μg) per mL of sample tested

  • Representative Standard Curve using ab108856.

Protocols

References

This product has been referenced in:

  • Tran TTT  et al. APOE genotype influences the gut microbiome structure and function in humans and mice: relevance for Alzheimer's disease pathophysiology. FASEB J N/A:fj201900071R (2019). Read more (PubMed: 30958695) »
  • Lv LL  et al. Effect of haptoglobin on the treatment of chronic hepatitis B with interferon. Exp Ther Med 18:1417-1425 (2019). Read more (PubMed: 31316628) »
See all 6 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Question
Answer

You do not need to subtract a blank for competitive ELISA. If you do, you will end up with negative values.

Read More

Question
Answer

Thank you for your patience with this reply.

I heard back from the lab that for the kits below, the antibodies were raised against recombinant proteins. The epitopes have not been mapped.

1) Apolipoprotein A1 Human ELISA Kit
2) Apolipoprotein E Human ELISA Kit
3) Haptoglobin Human ELISA Kit
4) Hemopexin Human ELISA Kit
5) Kininogen Human ELISA Kit
6) Retinol binding Protein 4 Human ELISA Kit

I'm sorry that this information could not be more helpful. Please contact us with any other questions.

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Answer

Thank you for your inquiry.

I heard back from the lab that serum that has undergone hemolysis will have a small interference with the haptoglobulin, but you should be able to use the sample. You should do a background control in this case.

I hope this information helps. Please contact us with any other questions.

Read More

Answer

Thank you for contacting us and your interest in our products. I am sorry for the delay in getting back to you.
The Haptoglobulin Human ELISA Kits (ab108856 and ab108858) can be used in order to quantify the level of haptoglobulin in various biological samples. With conditioned culture media samples the kit ab108858 would be most appropriate to use. This has been used with cell culture supernatant, saliva, milk, and urine.
I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

Read More

Answer

Thank you very much for your interest in ab108856

To our knowledge, ab108856 has not been tested in Urine samples. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.

2. Our scientists will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

Read More

Answer

Thank you very much for your call last week and for your patience while I have been in touch with the lab regarding your enquiry.

I have received the following information from the lab:

The proper value for Haptoglobulin in human plasma/serum is 0.3-2 mg/ml

We suggest to store samples for 3 months at -20C. However, we tested samples stability and I think that your samples stored at -80C should work.
I hope that this information will be useful, but please let me know if you have any further questions or if there is anything else that we can do for you, and I'll be happy to help.

Read More

Answer

Te pido disculpas por la espera y agradezco tu paciencia.

Me confirman mis compañeros del laboratorio que ambos kits (ab108856 y ab108858) detectan la subunidad beta de la proteína. Además, si la sensibilidad del kit ab108856 os conviene más, no hay inconvenientes en usarlo en muestras de orina, pero en este tipo de muestra no lo hemos optimizado, por lo que tendréis que probar distintas condiciones para encontrar las más adecuadas a vuestro experimento concreto.

Espero haberte ayudado. Para cualquier otra consulta o sugerencia, por favor no dudes en volvernos a contactar.

Read More

Answer

Thank you for your reply.

The standard curve you had sent me originally looked good. I assume these are the same data.

I apologize, I was mistaken.The blank has the highest value, thus you would not be able to subtract it from the other values - you are right. The blank just serves as a 0 ug/ml reference point.

You would make the standard curve from the five points STD01-05.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your reply.

You can use percentages in your data analysis. It is fine to use zero concentration (blank) as 100% absorbance for competitive ELISA.

We have never used Masterplex so I do not know how it works. If it is a scientific software, your customer should be able to assign concentration to each standard OD point and this software should do all of calculations and provide him with a graph.

Please let me know if there is anything else I can help you with.

Read More

Answer

Thank you for your email.

Yes, in my previous email I had been refering to the section 6. Preparation of the reagents, page 7, of the protocol (https://www.abcam.com/ps/products/108/ab108856/documents/ab108856%20Haptoglobin%20Human%20ELISA%20Kit%20(Website).pdf):


3. Standard Curve: Reconstitute the Haptoglobulin Standard with the appropriate amount of Diluent to generate a solution of 40 μg/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare duplicate or triplicate standard points by serially diluting the standard solution (20 μg/ml) 1:2 with Diluent, then 1:4 to produce 20, 5, 1.25, 0.313, and 0.078 μg/ml solutions. Diluent serves as the zero standard (0 μg/ml).


But first, you would need to dilute the Diluent 1:10 with reagent grade water - as explained in step 2.

The diluted diluent is your blank which you would substract from your samples and standard curve. Since the standard curve is presented in a log-log scale, the zero-datapoint would not be includedin the curve.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

1-10 of 14 Abreviews or Q&A

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