PIIINP ELISA Kit

Produktnummer ABIN415073

Produktdetails PIIINP ELISA Kit

Target: PIIINP (Procollagen III N-Terminal Propeptide (PIIINP))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.62 ng/mL - 40 ng/mL

Untere Nachweisgrenze: 0.62 ng/mL

Applikation: ELISA

Verwendungszweck: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of PIIINP in Serum,Plasma,Biological Fluids

Proben: Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate

Analytische Methode: Quantitative

Spezifität:

This assay has high sensitivity and excellent specificity for detection of Procollagen III N-Terminal Propeptide (PIIINP).
No significant cross-reactivity or interference between Procollagen III N-Terminal Propeptide (PIIINP) and analogues was observed.

Kreuzreaktivität (Details): No significant cross-reactivity or interference between Procollagen III N-Terminal Propeptide (PIIINP) and analogues was observed.

Sensitivität: 0.241 ng/mL

Bestandteile:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Anwendungsinformationen

Applikationshinweise:

• Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
• The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
• Kits from different batches may be a little different in detection range, sensitivity and color developing time.
• Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
• Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
• There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
• Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
• Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
• Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
• Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.

Kommentare:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Probenmenge: 100 μL

Testdauer: 3 h

Plattentyp: Pre-coated

Protokoll: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Procollagen III N-Terminal Propeptide (PIIINP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Procollagen III N-Terminal Propeptide (PIIINP). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Procollagen III N-Terminal Propeptide (PIIINP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Procollagen III N-Terminal Propeptide (PIIINP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Aufbereitung der Reagenzien:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 160 ng/mL. Firstly dilute the stock solution to 40 ng/mL and the diluted standard serves as the highest standard (40 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 40 ng/mL, 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Testpräzision:

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Procollagen III N-Terminal Propeptide (PIIINP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Procollagen III N-Terminal Propeptide (PIIINP) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Handhabung: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Lagerung: 4 °C

Informationen zur Lagerung:

• For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
• For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
  Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
• For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.

Haltbarkeit: 6 months

Referenzen für PIIINP ELISA Kit (ABIN415073)

Mukherjee, Zhelnin, Sanfiz, Pan, Li, Yarde, McCarty, Jarai: "Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype." in: American journal of translational research, Vol. 11, Issue 3, pp. 1531-1540, (2019) (PubMed).

Meel, Nethononda, Libhaber, Dix-Peek, Peters, Essop: "Assessment of myocardial fibrosis by late gadolinium enhancement imaging and biomarkers of collagen metabolism in chronic rheumatic mitral regurgitation." in: Cardiovascular journal of Africa, Vol. 29, Issue 3, pp. 150-154, (2019) (PubMed).

Li, Zhang, Sun, Li, Yu, Fu, Chen: "Circulating matrix metalloproteinases and procollagen propeptides in inguinal hernia." in: Hernia : the journal of hernias and abdominal wall surgery, Vol. 22, Issue 3, pp. 541-547, (2019) (PubMed).

Rubiś, Wiśniowska-Smiałek, Wypasek, Rudnicka-Sosin, Hlawaty, Leśniak-Sobelga, Kostkiewicz, Podolec et al.: "12-month patterns of serum markers of collagen synthesis, transforming growth factor and connective tissue growth factor are similar in new-onset and chronic dilated cardiomyopathy in patients both ..." in: Cytokine, Vol. 96, pp. 217-227, (2018) (PubMed).

Abdul Alim, Domeij-Arverud, Nilsson, Edman, Ackermann: "Achilles tendon rupture healing is enhanced by intermittent pneumatic compression upregulating collagen type I synthesis." in: Knee surgery, sports traumatology, arthroscopy : official journal of the ESSKA, Vol. 26, Issue 7, pp. 2021-2029, (2018) (PubMed).

Liu, Chen, Zhen, Ho, Tsang, Yuen, Lam, Tse, Yiu: "Relationship of biomarkers of extracellular matrix with myocardial function in Type 2 diabetes mellitus." in: Biomarkers in medicine, Vol. 11, Issue 7, pp. 569-578, (2018) (PubMed).

Rubiś, Wiśniowska-Śmiałek, Dziewięcka, Rudnicka-Sosin, Kozanecki, Podolec: "Prognostic value of fibrosis-related markers in dilated cardiomyopathy: A link between osteopontin and cardiovascular events." in: Advances in medical sciences, Vol. 63, Issue 1, pp. 160-166, (2018) (PubMed).

Sunman, Canpolat, Yorgun, Özkan, Yalçın, Bayrak, Şahiner, Kaya, Özkara, Aytemir, Oto: "Association between reverse electrical remodeling and cardiac fibrosis markers in patients with cardiac resynchronization therapy." in: Turk Kardiyoloji Dernegi arsivi : Turk Kardiyoloji Derneginin yayin organidir, Vol. 46, Issue 2, pp. 84-91, (2018) (PubMed).

Jotwani, Katz, Ix, Gutiérrez, Bennett, Parikh, Cummings, Sarnak, Shlipak: "Urinary Biomarkers of Kidney Tubular Damage and Risk of Cardiovascular Disease and Mortality in Elders." in: American journal of kidney diseases : the official journal of the National Kidney Foundation, Vol. 72, Issue 2, pp. 205-213, (2018) (PubMed).

Yılmaz, Yıldırım, Baydar, Altun, Erdem: "Urinary Type III Procollagen Is Associated With Chronic Allograft Dysfunction and Predicts Graft Survival." in: Transplantation proceedings, Vol. 49, Issue 2, pp. 281-287, (2017) (PubMed).

Sokal, Lenarczyk, Kowalski, Mitrega, Pluta, Stabryla-Deska, Streb, Urbanik, Krzeminski, Kalarus: "Prognostic value of collagen turnover biomarkers in cardiac resynchronization therapy: A subanalysis of the TRUST CRT randomized trial population." in: Heart rhythm : the official journal of the Heart Rhythm Society, Vol. 13, Issue 5, pp. 1088-95, (2016) (PubMed).

Rubiś, Wiśniowska-Śmialek, Wypasek, Biernacka-Fijalkowska, Rudnicka-Sosin, Dziewiecka, Faltyn, Khachatryan, Karabinowska, Kozanecki, Tomkiewicz-Pająk, Podolec: "Fibrosis of extracellular matrix is related to the duration of the disease but is unrelated to the dynamics of collagen metabolism in dilated cardiomyopathy." in: Inflammation research : official journal of the European Histamine Research Society ... [et al.], Vol. 65, Issue 12, pp. 941-949, (2016) (PubMed).

Lin, Kuo, Lee, Chou, Chen, Yeh, Huang, Hung, Liu, Ho, Wu: "5-methoxytryptophan is a potential marker for post-myocardial infarction heart failure - a preliminary approach to clinical utility." in: International journal of cardiology, Vol. 222, pp. 895-900, (2016) (PubMed).

Rubiś, Wiśniowska-Śmiałek, Biernacka-Fijałkowska, Rudnicka-Sosin, Wypasek, Kozanecki, Dziewięcka, Faltyn, Karabinowska, Khachatryan, Hlawaty, Leśniak-Sobelga, Kostkiewicz, Płazak, Podolec: "Left ventricular reverse remodeling is not related to biopsy-detected extracellular matrix fibrosis and serum markers of fibrosis in dilated cardiomyopathy, regardless of the definition used for LVRR." in: Heart and vessels, Vol. 32, Issue 6, pp. 714-725, (2016) (PubMed).

Buko, Lukivskaya, Naruta, Belonovskaya, Tauschel: "Protective Effects of Norursodeoxycholic Acid Versus Ursodeoxycholic Acid on Thioacetamide-induced Rat Liver Fibrosis." in: Journal of clinical and experimental hepatology, Vol. 4, Issue 4, pp. 293-301, (2015) (PubMed).

Bielecka-Dabrowa, Michalska-Kasiczak, Gluba, Ahmed, Gerdts, von Haehling, Rysz, Banach: "Biomarkers and echocardiographic predictors of myocardial dysfunction in patients with hypertension." in: Scientific reports, Vol. 5, pp. 8916, (2015) (PubMed).

Bielecka-Dabrowa, Gluba-Brzózka, Michalska-Kasiczak, Misztal, Rysz, Banach: "The multi-biomarker approach for heart failure in patients with hypertension." in: International journal of molecular sciences, Vol. 16, Issue 5, pp. 10715-33, (2015) (PubMed).

Page, Charles, Piccinini, Nicolaidou, Taylor, Midwood: "Raised circulating tenascin-C in rheumatoid arthritis." in: Arthritis research & therapy, Vol. 14, Issue 6, pp. R260, (2014) (PubMed).

Durnea, Khashan, Kenny, Durnea, Smyth, OReilly: "Prevalence, etiology and risk factors of pelvic organ prolapse in premenopausal primiparous women." in: International urogynecology journal, Vol. 25, Issue 11, pp. 1463-70, (2014) (PubMed).

Banerjee, Mukherjee, Ghosh, Biswas, Dutta, Pattari, Chatterjee, Bandyopadhyay: "Clinical significance of markers of collagen metabolism in rheumatic mitral valve disease." in: PLoS ONE, Vol. 9, Issue 3, pp. e90527, (2014) (PubMed).

Details zu PIIINP

Target: PIIINP (Procollagen III N-Terminal Propeptide (PIIINP))

Andere Bezeichnung: PIIINP (PIIINP Produkte)

UniProt: P02461