Overview

  • Product name

    Human Apolipoprotein AI ELISA Kit (APOA1)
    See all Apolipoprotein A I 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

    1 µg/ml
  • Range

    0.625 µg/ml - 20 µg/ml
  • Recovery

    98 %

  • Assay time

    3h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Apolipoprotein AI Human (APOA1) in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Apolipoprotein AI levels in plasma and serum.


    An Apolipoprotein AI specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently biotinylated Apolipoprotein AI 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 Apolipoprotein AI captured in plate.


    Get results in 90 minutes with Human Apolipoprotein AI ELISA Kit (ab189576) from our SimpleStep ELISA® range.


    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

    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
    1X Biotinylated Human Apolipoprotein AI (Lyophilized) 1 vial
    Apolipoprotein AI Microplate (12 x 8 well strips) 1 unit
    Apolipoprotein AI Standard 1 vial
    Chromogen Substrate 1 x 8ml
    Sealing Tapes 3 units
    Stop Solution 1 x 12ml
  • Research areas

  • Function

    Participates in the reverse transport of cholesterol from tissues to the liver for excretion by promoting cholesterol efflux from tissues and by acting as a cofactor for the lecithin cholesterol acyltransferase (LCAT). As part of the SPAP complex, activates spermatozoa motility.
  • Tissue specificity

    Major protein of plasma HDL, also found in chylomicrons. Synthesized in the liver and small intestine. The oxidized form at Met-110 and Met-136 is increased in individuals with increased risk for coronary artery disease, such as in carrier of the eNOSa/b genotype and exposure to cigarette smoking. It is also present in increased levels in aortic lesions relative to native ApoA-I and increased levels are seen with increasing severity of disease.
  • Involvement in disease

    High density lipoprotein deficiency 2
    High density lipoprotein deficiency 1
    APOA1 mutations may be involved in the pathogenesis of amyloid polyneuropathy-nephropathy Iowa type, also known as amyloidosis van Allen type or familial amyloid polyneuropathy type III (PubMed:3142462 and PubMed:2123470). The clinical picture is dominated by neuropathy in the early stages of the disease and nephropathy late in the course. Death is due in most cases to renal amyloidosis.
    Amyloidosis 8
  • Sequence similarities

    Belongs to the apolipoprotein A1/A4/E family.
  • Post-translational
    modifications

    Palmitoylated.
    Met-110 and Met-136 are oxidized to methionine sulfoxides.
    Phosphorylation sites are present in the extracellular medium.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Apo-AI
    • ApoA I
    • ApoA-I
    • APOA1
    • APOA1_HUMAN
    • Apolipoprotein A 1
    • Apolipoprotein A-I
    • Apolipoprotein A-I(1-242)
    • Apolipoprotein A1
    • Apolipoprotein of high density lipoprotein
    • ApolipoproteinAI
    • Brp14
    • Ltw1
    • Lvtw1
    • MGC117399
    • Sep1
    • Sep2
    • Truncated apolipoprotein A-I
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab108804 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 concentration.

Images

  • Apolipoprotein A1 measured in biological fluids and cell culture medium with background signal subtracted (duplicates +/- SD).
  • Representative Standard Curve using ab108804

Protocols

References

This product has been referenced in:

  • Aguirre-Portolés C  et al. ABCA1 overexpression worsens colorectal cancer prognosis by facilitating tumour growth and caveolin-1-dependent invasiveness, and these effects can be ameliorated using the BET inhibitor apabetalone. Mol Oncol 12:1735-1752 (2018). Read more (PubMed: 30098223) »
  • Wang Y  et al. The mechanism of apoliprotein A1 down-regulated by Hepatitis B virus. Lipids Health Dis 15:64 (2016). Read more (PubMed: 27015844) »
See all 6 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Answer

Only the assays for APOB (ab108807) and APOE (ab108813) have been tested with CSF. They are suitable for this kind of sample. The APOA1 assays have not been tested with CSF yet, but there are plans to test ab108803. Kit ab108804, the competitive ELISA for APOA1, will not be tested with CSF, and it is not known if it is suitable for assays of CSF.

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Answer

Vielen Dank für Ihre Anfrage.


Leider haben wir kein weiteres Kit der Charge GR104136 mehr in unserem Lager (weltweit).

Diese Kits ist normalerweise sehr zuverlässig, aber Sie haben Recht, es gibt eine Komponente, die nie gleich ist: Der Standart in ist ein aufgereinigtes humanes Protein, das in keinem Lot die gleiche Konzentration aufweist. Das heißt, für jedes Lot muss eine neue Etablierung stattfinden, und daher kann es zu so starken Abweichungen zwischen den verschieden Lots kommen.


Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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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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Abreview for Apolipoprotein A1 Human ELISA Kit

Excellent Excellent 5/5 (Ease of Use)
Abreviews

Sample: Human Serum (Human plasma)
Specification: Human plasma
Amount of Sample: 2 µg
Type of Kit Used: ELISA
Positive control: recombinant apolipoprotein A1
Did you use the Abcam Kit protocol?: Yes

Abcam user community

Verified customer

Submitted Jan 15 2013

Answer


Below is the information for the standard specific to your lot:

Standard Curve: Reconstitute the 120 mg of Apo A-I Standard with 3 ml of Diluent to generate a solution of 40 mg/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 (40 mg/ml) 1:2 with equal volume Diluent to produce 20, 10, 5, 2.5, 1.25, and 0.625 mg/ml solutions. Diluent serves as the zero standard (0 mg/ml). Any remaining solution should be frozen at -200C.

I hope this information is helpful to you. Please do not hesitate to contact us with any further questions.

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Answer


I contacted the laboratory, and they responded that the Apo A-1 standard should be in micrograms, not nanograms. The current lot's standard is delivered at 60ug and 3mL of diluent would make the 20ug/ml solution. If you respond with your specific lot# or order# I can tell you what the standard size for your lot should have been.

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Answer

Vielen Dank für Ihre Anfrage. Ich entschuldige mich für die Wartezeit, ich musste auf nähere Angaben aus dem produzierenden Labor warten.

Ich freue mich Ihnen nun bestätigen zu können, dass beide Standards aus aufgereinigten humanen Proteinen bestehen.

Die Menge an Standard ist Lot-abhängig und kann auf dem Röhrchen gefunden werden.

Ich wünsche Ihnen viel Erfolg für Ihre Versuche und verbleibe

Read More

Answer

Thank you for contacting Abcam.

After receiving the customer's complaint and abreview showing cross-reactivity with apoB using the kit ab108804 in May, we retested the kit for cross-reactivity. There was no significant cross-reactivity with other Apo proteins in the retesting. We used pure proteins and tested cross-reactivity at high concentration (same concentration as one of the highest standard point). It was suspected that the customer may not have used pure apoB protein, potentially explaining the results.

The cross-reactivity testing results are given below:




Species
% Cross Reactivity

Canine
1%

Bovine
None

Monkey
<15% (Suggest dilution 1:20 for plasma/serum)

Mouse
1%

Rat
1%

Swine
1%

Rabbit
None

Proteins
% Cross Reactivity

Apo AII
<5%

Apo B
<5%

Apo CI
<5%

Apo CII
<5%

Apo CIII
<5%

Apo E
<5%

Apo H
<5%

Apo M
<5%

Apo AI
100%






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

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Answer


Here is the shelf life information you requested:
ab108804 - 6 months if kit is unused and reagents stored at 4 deg C or -20 deg C as appropriate; 1 month after opening and limiting reagent is microtiter plate storage
ab108807 - 6 months if kit is unused and reagents stored at 4 deg C or -20 deg C as appropriate; 1 month after opening and limiting reagents are microtiter plate and opened Diluent storage
ab108813- 6 months if kit is unused and reagents stored at 4 deg C or -20 deg C as appropriate; 1 month after opening and limiting reagents are microtiter plate and opened Diluent storage

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Answer

Thank you for your inquiry.
I am assuming that you are planning to use ab108804? https://www.abcam.com/index.html?datasheet=108804 https://www.abcam.com/index.html?datasheet=108804.
I can confirm that ab108804 has not been tested with cell culture supernatant yet.
I believe it should be suitable for this sample type two but this has to be determined experimentally by the user.
I strongly suggest to use serum (FCS) free media. I suspect that FCS will interfere with the correct measurement of human Apo A1.
I am sorry I could not be of more help and hope this information is nevertheless helpful.

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1-10 of 13 Abreviews or Q&A

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