製品の概要

  • 製品名

    Human Kininogen ELISA Kit (KNG)
    Kininogen キット 製品一覧
  • 検出方法

    Colorimetric
  • 再現性

    Intra-Assay(同時再現性)
    サンプル N 平均値 SD CV%
    Overall 5.6%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    Overall 10.1%
  • サンプルの種類

    Cell culture supernatant, Serum, Plasma
  • アッセイタイプ

    Competitive
  • 検出感度

    > 50 ng/ml
  • 検出範囲

    0.063 µg/ml - 4 µg/ml
  • 添加回収試験

    97 %

  • 全工程の試験時間

    3h 00m
  • ステップ

    Multiple steps standard assay
  • 種交差性

    交差種: Human
  • 製品の概要

    Abcam’s Kininogen (KNG) Human in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Kininogen in plasma and serum samples.


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


    Get results in 90 minutes with Human HMW Kininogen ELISA Kit (ab189574) 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.

  • アプリケーション

    適用あり: Competitive ELISAmore details
  • 試験プラットフォーム

    Microplate

製品の特性

  • 保存方法

    Please refer to protocols.
  • 内容 1 x 96 tests
    100X Streptavidin-Peroxidase Conjugate 1 x 80µl
    10X Diluent N Concentrate 1 x 30ml
    1X Biotinylated Human Kininogen (Lyophilized) 2 vials
    20X Wash Buffer Concentrate 1 x 30ml
    Chromogen Substrate 1 x 8ml
    Kininogen Microplate (12 x 8 well strips) 1 unit
    Kininogen Standard 2 vials
    Sealing Tapes 3 units
    Stop Solution 1 x 12ml
  • 研究分野

  • 機能

    (1) Kininogens are inhibitors of thiol proteases; (2) HMW-kininogen plays an important role in blood coagulation by helping to position optimally prekallikrein and factor XI next to factor XII; (3) HMW-kininogen inhibits the thrombin- and plasmin-induced aggregation of thrombocytes; (4) the active peptide bradykinin that is released from HMW-kininogen shows a variety of physiological effects: (4A) influence in smooth muscle contraction, (4B) induction of hypotension, (4C) natriuresis and diuresis, (4D) decrease in blood glucose level, (4E) it is a mediator of inflammation and causes (4E1) increase in vascular permeability, (4E2) stimulation of nociceptors (4E3) release of other mediators of inflammation (e.g. prostaglandins), (4F) it has a cardioprotective effect (directly via bradykinin action, indirectly via endothelium-derived relaxing factor action); (5) LMW-kininogen inhibits the aggregation of thrombocytes; (6) LMW-kininogen is in contrast to HMW-kininogen not involved in blood clotting.
  • 組織特異性

    Secreted in plasma. T-kinin is detected in malignant ovarian, colon and breast carcinomas, but not in benign tumors.
  • 関連疾患

    Defects in KNG1 are the cause of high molecular weight kininogen deficiency (HMWK deficiency) [MIM:228960]. HMWK deficiency is an autosomal recessive coagulation defect. Patients with HWMK deficiency do not have a hemorrhagic tendency, but they exhibit abnormal surface-mediated activation of fibrinolysis.
  • 配列類似性

    Contains 3 cystatin domains.
  • 翻訳後修飾

    Bradykinin is released from kininogen by plasma kallikrein.
    Hydroxylation of Pro-383 occurs prior to the release of bradykinin.
    Phosphorylation sites are present in the extracelllular medium.
    N- and O-glycosylated. O-glycosylated with core 1 or possibly core 8 glycans.
  • 細胞内局在

    Secreted > extracellular space.
  • Information by UniProt
  • 別名

    • Alpha-2-thiol proteinase inhibitor
    • BDK
    • BK
    • BK, included
    • bradykinin
    • bradykinin, included
    • Fitzgerald factor
    • Fitzgerald factor, included
    • Flaujeac factor, included
    • High molecular weight kininogen
    • High molecular weight kininogen, included
    • HMWK
    • HMWK, included
    • Ile-Ser-Bradykinin
    • Kallidin I
    • Kallidin II
    • kininogen 1
    • KNG
    • KNG1
    • KNG1_HUMAN
    • LMWK, included
    • Low molecular weight growth-promoting factor
    • Low molecular weight kininogen, included
    • Williams factor, included
    • Williams-Fitzgerald-Flaujeac factor
    see all
  • 参照データベース

関連製品

アプリケーション

Our Abpromise guarantee covers the use of ab108875 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Competitive ELISA Use at an assay dependent concentration.

画像

  • Kininogen measured in biological fluids showing quantity (ng) per mL of tested sample. Samples diluted 400-800 fold.

  • Representative Standard Curve using ab108875.

プロトコール

参考文献

This product has been referenced in:

  • Zhang Y  et al. Application of Isobaric Tags for Relative and Absolute Quantification (iTRAQ) Coupled with Two-Dimensional Liquid Chromatography/Tandem Mass Spectrometry in Quantitative Proteomic Analysis for Discovery of Serum Biomarkers for Idiopathic Pulmonary Fibrosis. Med Sci Monit 24:4146-4153 (2018). Read more (PubMed: 29909421) »
  • Liu C  et al. Serum Protein KNG1, APOC3, and PON1 as Potential Biomarkers for Yin-Deficiency-Heat Syndrome. Evid Based Complement Alternat Med 2016:5176731 (2016). Read more (PubMed: 27843478) »
See all 2 Publications for this product

レビューと Q&A

1-10 of 14 Abreviews or Q&A

Kininogen (KNG) Human ELISA Kit

Excellent Good 4/5 (Ease of Use)
Abreviews
Sample: EDTA human plasma stored at -80ºC
Amount of Sample: 3 ul
Type of Kit Used: ELISA
Manufacturer instructions were followed.
Additional data
We diluted 3 ul of plasma in 600 ul of 1XDiluent N (1:200)
Washings were done on a Microplate Washer machine with 300 ul of wash buffer 1X.
The standards data generated an excellent curve.
All samples assayed showed data inside the curve.

Abcam user community

Verified customer

投稿 Dec 07 2015

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

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

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Answer

Thank you for your reply.

I believe bulk discounts begin at 5 items per order. For a specific quote, please send your request to mailto:sales@abcam.com and one of our Sales Representatives will be able to help you with discounts.

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

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Question
Answer

Thank you for your reply.

The source of this kit is considered proprietary, so I apologize that we are not able to provide this information.

They did release that the antigen for the antibodies in the kitis a full length protein Kininogen (MW 120,000) purified from Human source and with greater than 98% purity. But again, these are polyclonal antibodies, so they will have multiple binding epitopes.

The lab does not know of any publications using this kit, but they provided these background references for you:

(1) Reddigari SR et al. (1993) Blood 81:1306-1311
(2) Maier M et al. (1983) Proc. Natl. Acad. Sci. USA 80:3928-3932
(3) Kellermann J et al. (1986) Eur. J. Biochem. 154:471-478
(4) Kitamura N et al. (1985) J. Biol. Chem. 260:8610-8617
(5) Kerbiriou DM and Griffin JH (1979) J. Biol. Chem. 254:12020-12027
(6) Coffman LG et al. (2009) Proc. Natl. Acad. Sci. USA 106:570-575
(7) Hassan S et al. (2007) Am. J. Physiol Heart Circ Physiol. 292:H2959-H2965

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

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Answer

Thank you for your inquiry.

The two antibodies in this kit are polyclonal and therefore will have multiple binding epitopes. The supplying lab does not have any information about the immunogen details for these antibodies.

I am 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 email.

The antibody in the kit was raised against full length Kiniogen. It will react to multiple epitopes in the protein.

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

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Answer

Thank you for contacting us.

Kate is currently away from the office but I will try to help as best I can with your enquiry. Firstly I am sorry for the delay in getting back to you. If the lot number of the kit your customer has been using is of the lot number GR86524 then a total of 32 μg of Human Kininogen Standard is provided with the kit.

The protocol describes making a solution of 8 μg/mL initially, then dilution further to measure the standard curve. To make the 8 μg/mL Human Kininogen Standard stock solution the customer will need to reconstitute the lypholised product in 4 mL of Diluent.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

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Answer

I would like to thank you for noticing this and bringing it to our attention.

It appears in this case that there has been a mislabeled reagent for this kit. Our "Biotinylated Human Kininogen Antibody: 1 vial, lyophilized" should read "Biotinylated Human Kininogen 1 vial, lyophilized".

I have made to appropriate group aware of this error and hope that this will be remedied on the website, manual and kit components soon.

Thank you again. I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

I would like to encourage you to take advantage of our Abreview program. It takes just a few minutes to leave a review and you can collect Abpoints which you may redeem for Abcam products or Amazon gift cards while at the same time sharing information about the product with your colleagues worldwide. More information may be at:

https://www.abcam.com/index.html?pageconfig=resource&rid=10332

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Question

Hello, I would like clarification regarding how your kininogen ELISA kit for plasma and serum samples (ab108875) works. There seems to be conflicting information on whether it is a sandwich ELISA or competitive ELISA. The Datasheet tab on its webpage https://www.abcam.com/Kininogen-Human-ELISA-Kit-ab108875.html says it is a sandwich ELISA. However, the protocol on page 2 says "This assay employs a quantitative competitive enzyme immunoassay technique", but yet it then goes on to say that the plate is pre-coated with anti-kininogen antibody (as in a sandwich ELISA), and that kininogen in samples is "sandwiched" by the immobilized primary antibody and the biotinylated primary anti-kininogen antibody that you add to the plate after your standards and samples. It is clear from the standard curve on p.10 (and I have done the assay and my results agree) that in fact this is a competitive ELISA, in that the absorbance goes down as the kininogen concentration in the standard goes up. However, in that case it does not make sense to me that the plate would be coated with primary anti-kininogen antibody. If it were, there would be no competition-- the more kininogen in your sample, the more would bind to the immobilized antibody, and the more biotinylated antibody could bind to that, which ultimately gives the color signal. Other competitive ELISAs I've seen work by either coating the plate with secondary antibody, and then you add primary antibody, sample, and biotinylated protein of interest (e.g. kininogen) to compete with the sample for the primary antibody (this is not the case here, as there is no biotinylated kininogen), or by coating the plate with the protein of interest, and thus the protein in your sample has to compete with the protein on the plate for the biotinylated primary antibody, and the more protein in your sample, the more biotinylated primary antibody gets washed away (thus reducing signal). So, is it possible that for this assay, the plate is actually coated with kininogen protein, as opposed to anti-kininogen antibody? I see that your other kininogen ELISA kit, for milk, urine, saliva, and cell culture samples, is truly a sandwich ELISA (with a curve showing the absorbance increases with concentration), so perhaps this is the source of the confusion? Thanks for your help with this,

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Answer

Thank you for contacting us regarding this. I have been in touch with the lab to try and clear this up.

This product is a competitive ELISA assay. The important thing is that Kininogen in your sample or in the standard is not biotinylated. On the other hand, we provide biotinylated-kininogen so there is a competition between biotinylated-kininogen and kininogen for binding to immobilized antibody on the plate.

I would like to encourage you to take advantage of our Abreview program. It takes just a few minutes to leave a review and you can collect Abpoints which you may redeem for Abcam products or Amazon gift cards while at the same time sharing information about the product with your colleagues worldwide. More information may be at:

https://www.abcam.com/index.html?pageconfig=resource&rid=10332

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.



Principle of the assay:

The Human Kininogen ELISA (Enzyme-Linked Immunosorbent Assay) kit, ab108875 is designed for detection of human Kininogen in plasma and serum. This assay employs a quantitative competitive enzyme immunoassay technique that measures human Kininogen in less than 3 hours. A polyclonal antibody specific for human Kininogen has been pre-coated onto a 96-well microplate with removable strips. Kininogen in standards and samples is competing with a biotinylated Kininigen sandwiched by the immobilized antibody and streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.

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