Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online

Creatinine Assay Kit (ab65340)

Reviews (2)Q&A (52)References (148)
Functional Studies - Creatinine Assay Kit (ab65340)
  • Functional Studies - Creatinine Assay Kit (ab65340)
  • Functional Studies - Creatinine Assay Kit (ab65340)
  • Functional Studies - Creatinine Assay Kit (ab65340)
  • Functional Studies - Creatinine Assay Kit (ab65340)
  • Immunodiffusion - Creatinine Assay Kit (ab65340)

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric/Fluorometric
  • Platform: Microplate reader
  • Assay time: 1 hr
  • Sample type: Other biological fluids, Serum, Urine

製品の概要

  • 製品名

    Creatinine Assay Kit
    Creatinine キット 製品一覧

  • 検出方法

    Colorimetric/Fluorometric

  • サンプルの種類

    Urine, Serum, Other biological fluids

  • アッセイタイプ

    Quantitative

  • 全工程の試験時間

    1h 00m

  • 製品の概要

    Creatinine Assay Kit ab65340 provides an accurate, convenient measure of creatinine concentration in biological fluids such as serum, urine or CSF.


    How the assay works
    In the creatinine assay protocol, creatinine is converted to creatine by creatininase, creatine is converted to sarcosine by creatinase. Sarcosine is then oxidized by sarcosine oxidase producing hydrogen peroxide. Hydrogen peroxide is then processed by a peroxidase causing a reaction with a probe to generate red color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm).


    Creatinine assay protocol summary


    - Add samples and standards to wells.
    - Add reaction mix and incubate for 60 min at 37°C.
    - Analyze with microplate reader.


    Getting the best performance from ab65340
    Unlike picric acid assays, this kit is suitable for serum creatinine determinations, as well as for urine and other biological samples.


    For deproteinization of samples: Better results are typically seen with this assay when using a 10kda filter for sample deproteinization than when using the PCA method.


    Related and recommended products
    Creatinine assay kit ab65340 is commonly used by researchers to measure kidney function.


    Kidney function marker assays include:


    - Creatinine Assay Kit ab65340 - suitable for use as a serum creatinine assay and a urine creatinine assay.
    - Mouse Albumin ELISA Kit ab108792 and Human Albumin ELISA Kit ab227933 – suitable for use with serum/plasma and urine
    - Albumin (BCG) Assay Kit (Colorimetric) ab235628 – suitable for use with serum
    - Urea Assay Kit ab83362 – suitable for use with serum/plasma and urine, and used as a blood urea nitrogen assay.


    The albumin-creatinine ratio is often measured by using in combination Creatine Assay Kit ab65340, and either Mouse Albumin ELISA Kit ab108792 or Human Albumin ELISA Kit ab227933.


    The urea-creatinine ratio is often measured by using in combination Urea Assay Kit ab83362 and Creatinine Assay Kit ab65340.


    Albumin Creatinine Ratio Assay Kit ab241018 is suitable for urine and serum, and combines a fluorescent albumin assay with a colorimetric creatinine assay in a convenient pack.


    Creatinine assay methods
    There are two established methods of creatinine assays:


    - Jaffe assay
    The Jaffe assay method is based on the reaction of creatinine with picric acid at an alkaline pH. The creatinine-picrate complex has an absorbance of 500 nm.


    - Enzymatic creatinine assay
    The standard enzymatic creatinine assay uses the method described above for ab65340.


    The Jaffe method is substantially simpler and cheaper than enzymatic creatinine assays, however it suffers substantially from interference by metabolites found in urine and serum.


    Other notes
    This product is manufactured by BioVision, an Abcam company and was previously called K625 Creatinine Colorimetric/Fluorometric Assay Kit. K625-100 is the same size as the 100 test size of ab65340.

  • 特記事項

    The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.

  • 試験プラットフォーム

    Microplate reader

製品の特性

関連製品

画像

  • Functional Studies - Creatinine Assay Kit (ab65340)
    Functional Studies - Creatinine Assay Kit (ab65340)Gai, Zhibo et al., Scientific reports vol. 7,1 9815., Fig 2, doi:10.1038/s41598-017-10168-6
    Mice were injected i.p. with 6-ECDCA before I/R injury. At 24 h after I/R, kidney, serum and urine samples were collected for measurements of creatinine in serum with ab65340
  • Functional Studies - Creatinine Assay Kit (ab65340)
    Functional Studies - Creatinine Assay Kit (ab65340)

    Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Functional Studies - Creatinine Assay Kit (ab65340)
    Functional Studies - Creatinine Assay Kit (ab65340)

    Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Functional Studies - Creatinine Assay Kit (ab65340)
    Functional Studies - Creatinine Assay Kit (ab65340)

    Creatinine measured in mouse and human serum plotted against RFU. Samples were diluted 5-10 fold.

  • Functional Studies - Creatinine Assay Kit (ab65340)
    Functional Studies - Creatinine Assay Kit (ab65340)

    Creatinine measured in mouse and human urine plotted against RFU. Samples were diluted 400-800 fold.

  • Immunodiffusion - Creatinine Assay Kit (ab65340)
    Immunodiffusion - Creatinine Assay Kit (ab65340)

    Creatinine levels in filtered human urine was measured in the presence or absence of creatininase (background signal subtracted.

参考文献 (148)

ab65340 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab65340 は 148 報の論文で使用されています。

  • Tham MS  et al. Deletion of Aurora kinase A prevents the development of polycystic kidney disease in mice. Nat Commun 15:371 (2024). PubMed: 38191531
  • Liao JT  et al. D-Limonene Promotes Anti-Obesity in 3T3-L1 Adipocytes and High-Calorie Diet-Induced Obese Rats by Activating the AMPK Signaling Pathway. Nutrients 15:N/A (2023). PubMed: 36678138
  • De Castro L  et al. Renal toxicity and biokinetics models after repeated uranium instillation. Sci Rep 13:4111 (2023). PubMed: 36914734
  • Alhelf M  et al. Prognostic significance of the genetic variant of lymphotoxin alpha (p.Thr60Asn) in egyptian patients with advanced hepatocellular carcinoma. Mol Biol Rep 50:4317-4327 (2023). PubMed: 36929286
  • Buchalter DB  et al. Systemic glucose-insulin-potassium reduces skeletal muscle injury, kidney injury, and pain in a murine ischaemia-reperfusion model. Bone Joint Res 12:212-218 (2023). PubMed: 37051831
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-10 of 54 Abreviews or Q&A

assay kit

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
perfect and easy to use for manips
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.

Abcam user community

Verified customer

投稿 Jan 24 2018

Creatinine in porcine urine samples

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Tested the kit in porcine urine samples and after deproteinisation, the kit worked. We used the colourimetric assay as per protocol in the booklet.
We used the deproteinisation protocol with PCA and KOH as provided in the booklet that accompanies the kit.
Had to dilute the deproteinised samples 50x to bring samples into the expected detectable range.
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.

Abcam user community

Verified customer

投稿 Jul 28 2015

Question


One my question is: if the samples of plasma and urine had been frozen and stored at -80oC, can this ab65340 Creatinine Assay Kit assay the creatinine value precisely and sensitively. or the samples could be frozen before assay?

Thank you very much!

Read More
Answer


I can confirm that frozen samples can be used for accurate results with this kit.

However, the success of the storage and quality of the sample will depend on multiple criteria. This would including the length of storage, the number of rounds the sample has gone through freeze-thaw, the time after which the samples were frozen after isolation etc. If you need to freeze the samples the best procedure in such case would be samples frozen immediately after their isolation, and stored without any free-thaw for a relative short period of time.

Read More

Question

Is this kit suitable for measurement of serum creatinine from rat samples

Read More
Answer

Creatine is a non-protein amino acid and therefore exists in the same form in all animals.

This Creatinine Assay Kit (ab65340) is therefore species independent which means that it can be used with all and any species which have creatinine. This kit can therefore be used on rat samples.

Read More

Question

Can this kit be used on samples collected in EDTA?

Read More
Answer

I have contacted the lab who have confirmed that they recommend Heparin over EDTA when preparing blood samples for use with this kit.

Read More

Question

I was hoping you could help me with a query regarding the Creatinine Assay Kit (ab65340). Does a background mix i.e assay buffer, creatinase, enzyme mix, probe have to be performed for the creatinine standards (aswell as for samples)?

Many thanks,

Read More
Answer

Please add background mix to standard and samples in separate wells? We recomend performing this in duplicate and then subtracting the mean from the original reading of standard and sample.

Read More

Question

I have run one plate but had many samples read above the highest standard using the fluorometric method.
Under step 1b) of the protocol it recommends diluting the probe 10x with DMSO which I didn't do.
However, under step 3) of the protocol it recommends using 0.4ul of the probe/well rather than 2ul which I did do.

My question is do I need to be doing both these dilutions for more accurate readings?

ie that would be essentially a 1/50 dilution of the probe/well.

Read More
Answer

We recommend diluting the probe 10x and then using 0.4ul of the probe for the best results for the fluorometric assay. Typically this works the best. This assay is very sensitive and having too much probe will result in very high RFU values beyond the range of the std. curve and can saturate the detector in the instrument.



Read More

Question

Compatible with glucose, acetone, acetate, acetic acid, etc. that typically interfere with GFR measurement?

Read More
Answer

We have not tested this kit individually in higher than physiological amounts of the moieties you mentioned. In typical samples, to our knowledge sarcosine and creatine can interfere with the assay which is resolved by running appropriate controls. Since the assay does not rely on the physical rate of filtration, these moieties are not likely to cause any issues.

Read More

Question


Estoy interesada en un kit de creatinina: Creatinine Assay Kit (ab65340).
Me gustaría saber para qué especies está diseñado?, cuál es su límite de
detección?cuánta cantidad de muestra se necesita? cuánto vale y cuánto
tardarían en mandarlo?

Read More
Answer

Gracias por contactarnos.
Este kit esta diseñado para mamíferos, y su límite de detección vendrá determinado por el valor más bajo de la lectura siempre que caiga dentro del rango linear de la recta estándar. Por ejemplo, el suero contiene entre 45-110 pmol/μl de creatinina.
Para muestras en las que no se conoce la cantidad aproximada de creatinina, sugerimos probar varias diluciones para asegurar que las lecturas caen dentro de los límites de la recta estándar.
En cuanto a la cantidad de muestra que se necesita, tal y como describe el protocolo, se requieren entre 0-50ul de muestra por pocillo.
El kit tiene un precio de 370€, y siendo que está en stock tarda entre uno y dos días laborables en llegar.

Read More

Question

uses mouse plasma (20 ul)
good standard curve

values of creatinine obtained is 10x higher than what is reported in literature
uses 2.5x dilution
obtained 2.2 mg/dL = 200 pmol / uL (Note in protocol: Serum contains ˜45-110 pmol/μl of creatinine.)
(literature based on HPLC methods states: 0.2 mg/dL as normal)

detects at 570 nm absorption

tips: consider deproteinization
check and substract creatine/sarcosine background
lab: creatininase: lot-to-lot variation in activity?
Note: Serum contains ˜45-110 pmol/μl of creatinine. need reference and for what kind of serum this refers to

Read More
Answer


I have contacted the lab for additional advice.

My suggestions -as we discuss over the phone - would be:
1) check the creatine and sarcosine background
2) use serum instead of plasma
3) deproteinize the samples as recommended in the protocol
4) In addition to what we discussed there can be an inherent difference in the methods used (HPLC vs. ELISA) which could account for the difference in values you might see after you substract the background and deproteinize.

If you check for literature values, I would suggest to look for other ELISA data and see what is reported for this methodology.

Read More

1-10 of 54 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com