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Hydroxyproline Assay试剂盒(Colorimetric) (ab222941)

Reviews (4) Submit a question References (26)
Hydroxyproline Assay Kit (Colorimetric) (ab222941)
  • Hydroxyproline Assay Kit (Colorimetric) (ab222941)

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 3 hr
  • Sample type: Other biological fluids, Serum, Tissue Homogenate, Urine

概述

  • 产品名称

    Hydroxyproline Assay试剂盒(Colorimetric)

  • 检测方法

    Colorimetric

  • 样品类型

    Urine, Serum, Other biological fluids, Tissue Homogenate

  • 检测类型

    Quantitative

  • 范围

    0.1 µg - 1 µg

  • 检测时间

    3h 00m

  • 种属反应性

    与反应: Mammals, Other species

  • 产品概述

    Hydroxyproline Assay Kit (Colorimetric) ab222941 provides a quick and convenient method to quantify hydroxyproline in tissue lysates and biological fluids such as urine and serum.


    The classical hydroxyproline assay protocol is based on the oxidation of hydroxyproline to a pyrrole intermediate followed by reaction with Ehrlich’s reagent dissolved in concentrated perchloric acid. Perchloric acid is a hazardous material that is both toxic and highly reactive, requiring special handling and waste-disposal protocols.


    This hydroxyproline assay protocol employs a proprietary acidic developer solution to accurately measure hydroxyproline in hydrolysates without the use of hazardous perchlorates. It is a quick and convenient protocol where hydroxyproline gets oxidized to form a reaction intermediate, which further in reaction forms brightly-colored chromophore that can be easily detected at OD 560 nm.


    The assay can detect as low as 0.05 µg hydroxyproline/well.

  • 说明

    This product is a replacement for Hydroxyproline Assay Kit K555; this kit does not contain the hazardous Perchloric acid which is present in K555.

    The components in this product are exactly the same as in K555, except that the Perchloric acid/Isopropanol Solution component in K555 has been replaced with the Developer component.

    If you would prefer to continue to use a kit in the format of K555, then use this product and:

    For safety reasons, Perchloric Acid will not be shipped by Abcam in the same shipment as either Isopropanol or this product.

    1) In a fume cupboard, mix 1 ml of 70% Perchloric Acid and 5 ml of Isopropanol to produce the Perchloric acid/Isopropanol Solution component previously provided in K555.

    2) Then follow the protocol for K555 available on www.biovision.com/documentation/datasheets/K555.pdf, using the components from this product, and the Perchloric acid/Isopropanol Solution that you produced.

    This product is manufactured by BioVision, an Abcam company and was previously called K226 Hydroxyproline Assay Kit (Perchlorate-Free). K226-100 is the same size as the 100 test size of ab222941.

    Hydroxyproline assay protocol summary:
    - add 10 N concentrated NaOH to samples and hydrolyze at 120ºC for 1 hr
    - cool on ice
    - neutralize with 10 N concentrated HCl, centrifuge and collect supernatant
    - add samples and standards to wells
    - evaporate wells to dryness by heating at 65ºC
    - add oxidation reagent mix to dissolve crystalline residue, and incubate at room temp for 20 min
    - add developer and incubate at 37ºC for 5 min
    - add DMAB concentrate and incubate for 45 min at 65ºC
    - analyze with microplate reader

    We are currently not able to offer K555 Hydroxyproline Colorimetric Assay Kit from BioVision and apologize for the inconvenience that this may cause. We recommend ab222941 as an alternative.

  • 平台

    Microplate reader

性能

  • 存放说明

    Store at -20°C. Please refer to protocols.
  • 组件 100 tests
    Chloramine T Concentrate 1 x 600µl
    Developer Solution I 1 x 5ml
    DMAB Concentrate 1 x 5ml
    Hydroxyproline Standard  1 x 100µl
    Oxidation Buffer 1 x 10ml
    Microplate Sealing Film 1 unit

  • 研究领域

  • 相关性

    Hydroxyproline, a non-essential amino acid derived from proline, with no known therapeutic use. Hydroxproline is used as a major component of structural protiens such as collagen, connective tissues, plant cell walls, tendons and ligaments and provides skin elasticity. Vitiman C is required for the conversion process from proline to hydroxyproline, a deficincy in vitiman C can lead to defects in collagen synthesis, thus, resulting in easy bruising, internal bleeding, breakdown of connective tissue of the ligaments and tendons, and increased risk to blood vessel damage. An unusual feature of this amino acid is that, it is not incorporated into collagen during biosynthesis at the ribosomal level, but is formed from proline by a posttranslational modification by an enzymatic hydroxylation reaction.

图片

  • Hydroxyproline Assay Kit (Colorimetric) (ab222941)
    Hydroxyproline Assay Kit (Colorimetric) (ab222941)
  • Hydroxyproline Assay Kit (Colorimetric) (ab222941)
    Hydroxyproline Assay Kit (Colorimetric) (ab222941)

    Typical Hydroxyproline standard calibration curve.

文献 (26)

发表研究结果有使用 ab222941？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab222941 被引用在 26 文献中.

  • Tang W  et al. METTL3 enhances NSD2 mRNA stability to reduce renal impairment and interstitial fibrosis in mice with diabetic nephropathy. BMC Nephrol 23:124 (2022). WB ; Mouse . PubMed: 35354439
  • He X  et al. Myofibroblast YAP/TAZ activation is a key step in organ fibrogenesis. JCI Insight 7:N/A (2022). PubMed: 35191398
  • Son YJ  et al. Yellow loosestrife (Lysimachia vulgaris var. davurica) ameliorates liver fibrosis in db/db mice with methionine- and choline-deficient diet-induced nonalcoholic steatohepatitis. BMC Complement Med Ther 21:44 (2021). PubMed: 33494735
  • Torres Y  et al. Cell-assembled extracellular matrix (CAM) sheet production: Translation from using human to large animal cells. J Tissue Eng 12:2041731420978327 (2021). PubMed: 33633827
  • Yu H  et al. Strontium ranelate promotes chondrogenesis through inhibition of the Wnt/ß-catenin pathway. Stem Cell Res Ther 12:296 (2021). PubMed: 34016181
View all Publications for this product

客户评价及客户问答

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1-4 of 4 Abreviews or Q&A

Porcine bone extracellular matrix quantification

 Inconclusive Excellent 5/5 (Ease of Use)
Abreviews
Overall, the kit is very easy to use with a clear protocol.
My sample is lyophilized porcine bone extracellular matrix hence the sample was dry and required a preliminary step in order to perform the protocol. I followed the instructions for tissue lysates and took different amounts of ECM (10, 100 mg). Next, I tried different heating times with NaOH (1 or 2 hrs). Additionally, I performed the protocol on cardiac ECM for comparison.
In all the different attempts the ECM didn't fully dissolve, in some samples, there were still residues and in others, there was visible fat. Hence, the results of the protocol were Inconclusive and additional work should be done in order to develop a reliable dissolving protocol.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MRS. Rotem Hayam

Verified customer

提交于 Oct 18 2021

Compatilbility Issue

 Below average Average 3/5 (Ease of Use)
Abreviews
I was unable to use this kit to quantify hydroxyproline in my mouse sarcoma cell lines. The kit only has one plate sealing film which was inconvenient. Additionally, I was only able to detect the standards. All of my sample wells only had background. I tried adding ascorbate salt to my cells prior to harvesting, and I was still unable to detect hydroxyproline despite having increased collagen transcript and protein (immunoblotting) in my samples.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MRS. Rebecca Dodd

Verified customer

提交于 Apr 09 2021

Straightfoward method to detect HYP - but not working on cell lysates!

 Good Good 4/5 (Ease of Use)
Abreviews
abreview image
I tried using this kit on 4T1 cell lysates. I incubated my cells with a P4HA1 inhibitor in order to see if i can detect any changes in HYP levels. unfortunately, HYP wasent detected even in the control samples. I used 10^6 cells with 100 DDW starting point but no almost no HYP was detected.
I wouldnt reccomend this kit for cells but it works for serum/media/tissue.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Yarden Ariav

Verified customer

提交于 Jul 13 2020

Absorption of collagen hydrolysate through zebrafish gills

 Good Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Collagen hydrolysate (CH) is a water-soluble substance, which together with the ability of fish to absorb compounds from the water through the gills [1], should allow its penetration. We tested the possibility of administering the drug to fish through water for absorption into the bloodstream. The fish were treated daily for a week with CH compound in three different doses: 10, 20 and 40 mg/L, compared with a control group (without CH). To assess the amount of CH in the serum, blood was extracted and hydroxyproline (HYP) was detected by using Hydroxyproline Assay Kit according to manufacturer’s protocol.
The values obtained were lower than the values of the calibration curve, these results can be explained by two possible reasons, it is possible that the material is not absorbed by the fish. Another possibility is that the values are lower than the identification threshold of the kit.

Rubinstein, A.L., Zebrafish assays for drug toxicity screening. Expert Opinion on Drug Metabolism & Toxicology, 2006. 2(2): p. 231-240.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Ms. nili vasserman

Verified customer

提交于 Jun 21 2019

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