CRP ELISA Kit

Produktnummer ABIN624960

Produktdetails CRP ELISA Kit

Target: CRP (C-Reactive Protein (CRP))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 2-600 pg/mL

Untere Nachweisgrenze: 2 pg/mL

Applikation: ELISA

Verwendungszweck: Human CRP (C-Reactive Protein) ELISA Kit for cell culture supernatants, plasma, and serum samples.

Proben: Plasma, Cell Culture Supernatant, Serum

Analytische Methode: Quantitative

Spezifität: This ELISA kit shows no cross-reactivity with the following cytokines tested: human Angiogenin, BDNF, BLC, ENA-78, FGF- 4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, GM-CSF, Leptin (OB), MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, MMP-1, -2, -3, -10, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensitivität: < 2 pg/mL

Produktmerkmale:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Bestandteile:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Benötigtes Material:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Anwendungsinformationen

Applikationshinweise: Recommended Dilution for serum and plasma samples20,000 - 200,000 fold

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Aufbereitung der Reagenzien:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample dilution: If your samples need to be diluted, 1x Assay Diluent D (Item K) should be used for dilution of serum/plasma/ culture supernatants/urine. Serum/plasma samples require around 20,000-200,000 fold dilution. For example, add 2 µL of serum/plasma into a tube with 398.0 µL 1x Assay Diluent D to prepare a 200-fold diluted sample. Mix through and then pipette 2 µL of prepared 200-fold diluted sample into a tube with 498 µL 1x Assay Diluent D to prepare a final 50,000 fold diluted sample. *Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
   3. Assay Diluent D (Item K) and Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use.
   4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Assay Diluent D (Item K) into Item C vial to prepare a 25 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 12 µL CRP standard from the vial of Item C, into a tube with 488 µL 1x Assay Diluent D to prepare a 600 pg/mL standard solution. Pipette 400myl 1x Assay Diluent D into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent D serves as the zero standard (0 pg/mL). 200 µL 12 µL standard + 488 µL 200myl 200 µL 200 µL 200 µL 600 200 66.67 22.22 7.407 2.469 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
   6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B (Item E) into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
   7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 300-fold with 1x Assay Diluent B (Item E). For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 40 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a final 300 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.

Testdurchführung:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
   2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
   3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
   4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
   5. Discard the solution. Repeat the wash as in step
   6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
   7. Discard the solution. Repeat the wash as in step
   8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
   9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Ergebnisberechnung:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Human CRP concentration (pg/mL) 1 10 100 1000 O D =4 50 n m 0.001 0.01 0.1 1 10 Assay Diluent D
Sensitivity: The minimum detectable dose of CRP is typically less than 2 pg/mL.
Recovery: Recovery was determined by spiking CRP into normal human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 116.7 106-127 Plasma 102.2 93-112 Cell culture media 106.2 95-116
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 107.7 113.1 84.76 Range ( %) 95-106 105-123 74-92 1:4 Average % of Expected 77.78 76.18 75.43 Range ( %) 70-88 70-87 68-87
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Testpräzision: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Haltbarkeit: 6 months

Referenzen für CRP ELISA Kit (ABIN624960)

Wang, Song, Chen, Yuan, Xu, Zhang, Tan, Yang, Yu, Lv: "The Long-Term Influence of Tissue Inhibitor of Matrix Metalloproteinase-1 in Patients with Mild to Moderate Coronary Artery Lesions in a Chinese Population: A 7-Year Follow-Up Study." in: Cardiology, Vol. 132, Issue 3, pp. 151-8, (2016) (PubMed).

Qian, Zhang, Guo, Ma, Chen, Wang, Zhao, Liu: "Polychlorinated biphenyls (PCBs) inhibit hepcidin expression through an estrogen-like effect associated with disordered systemic iron homeostasis." in: Chemical research in toxicology, Vol. 28, Issue 4, pp. 629-40, (2015) (PubMed).

Ciaffi, Cavassini, Genne, Delhumeau, Spycher Elbes, Hill, Wandeler, Fehr, Stoeckle, Schmid, Hirschel, Montecucco, Calmy: "Switch to etravirine for HIV-positive patients receiving statin treatment: a prospective study." in: European journal of clinical investigation, Vol. 45, Issue 7, pp. 720-30, (2015) (PubMed).

Verna, Patel, Bettencourt, Nguyen, Hernandez, Valasek, Kisselva, Brenner, Loomba: "Novel association between serum pentraxin-2 levels and advanced fibrosis in well-characterised patients with non-alcoholic fatty liver disease." in: Alimentary pharmacology & therapeutics, Vol. 42, Issue 5, pp. 582-90, (2015) (PubMed).

Zemer-Wassercug, Haim, Leshem-Lev, Orvin, Vaduganathan, Gutstein, Kadmon, Mager, Kornowski, Lev, Lev: "The effect of dabigatran and rivaroxaban on platelet reactivity and inflammatory markers." in: Journal of thrombosis and thrombolysis, Vol. 40, Issue 3, pp. 340-6, (2015) (PubMed).

Zhang, Wang, Miao, Chen, Qiang, Cui, Jing, Guo: "Magnetic ferroferric oxide nanoparticles induce vascular endothelial cell dysfunction and inflammation by disturbing autophagy." in: Journal of hazardous materials, Vol. 304, pp. 186-95, (2015) (PubMed).

Bil-Lula, Sochocka, Zato?ska, Szuba, Sawicki, Wo?niak: "Adenovirus type 9 enhances differentiation and decreases cytokine release from preadipocytes." in: Journal of medical virology, Vol. 87, Issue 2, pp. 230-9, (2014) (PubMed).

Rezvani, Gupta, Smith, Poursharifi, Marceau, Pérusse, Bouchard, Tchernof, Cianflone: "Cross-sectional associations of acylation stimulating protein (ASP) and adipose tissue gene expression with estradiol and progesterone in pre- and postmenopausal women." in: Clinical endocrinology, Vol. 81, Issue 5, pp. 736-45, (2014) (PubMed).

Bil-Lula, St?por, Krzywonos-Zawadzka, Wo?niak: "Is there any link between visceral obesity and adenovirus infections in the Polish population?" in: Advances in clinical and experimental medicine : official organ Wroclaw Medical University, Vol. 23, Issue 3, pp. 415-22, (2014) (PubMed).

Duan, Yu, Yu, Li, Wang, Geng, Jiang, Li, Zhou, Sun: "Silica nanoparticles induce autophagy and endothelial dysfunction via the PI3K/Akt/mTOR signaling pathway." in: International journal of nanomedicine, Vol. 9, pp. 5131-41, (2014) (PubMed).

Davies, Padlan, Sheriff: "Antibody-antigen complexes." in: Annual review of biochemistry, Vol. 59, pp. 439-73, (1990) (PubMed).

Details zu CRP

Target: CRP (C-Reactive Protein (CRP))

Andere Bezeichnung: C-Reactive Protein (CRP Produkte)

Synonyme: PTX1 ELISA Kit, crp ELISA Kit, AI255847 ELISA Kit, Aa1249 ELISA Kit, Ab1-341 ELISA Kit, Ab2-196 ELISA Kit, Ac1-114 ELISA Kit, Ac1262 ELISA Kit, Ac2-069 ELISA Kit, Ba2-693 ELISA Kit, APCS ELISA Kit, 0610010I23Rik ELISA Kit, AW743261 ELISA Kit, C77570 ELISA Kit, CRP2 ELISA Kit, CRP4 ELISA Kit, Crp ELISA Kit, ESP1 ELISA Kit, Hlp ELISA Kit, CRP5.1 ELISA Kit, zgc:152809 ELISA Kit, C-reactive protein ELISA Kit, C-reactive protein, pentraxin-related ELISA Kit, c-reactive protein, pentraxin-related ELISA Kit, cysteine rich protein 2 ELISA Kit, CRP ELISA Kit, crp ELISA Kit, Crp ELISA Kit, Crip2 ELISA Kit, LOC776376 ELISA Kit

Hintergrund: The Human CRP (C Reactive Protein) ELISA (Enzyme- Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human CRP in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human CRP coated on a 96-well plate. Standards and samples are pipetted into the wells and CRP present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human CRP antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of CRP bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.

Gen-ID: 1401

UniProt: P02741

Pathways: Carbohydrate Homeostasis