CRP ELISA Kit

Catalog No. ABIN955810

Product Details CRP ELISA Kit

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

Reactivity: Dog

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: The Dog High-Sensitive CRP ELISA is a highly sensitive two-site enzyme-linked immunoassay (ELISA) for the quantitative determination of C-reactive protein (CRP) in dog biological samples.

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Characteristics: In this assay the CRP present in the sample reacts with the anti-CRP antibodies that have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, anti-CRP antibody conjugated with horseradish peroxidase (HRP) is added. This HRP-conjugated antibody forms a complex with the previously bound CRP. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3',5,5'- tetramethylbenzidine (TMB). The quantity of bound enzyme is proportional to the concentration of CRP in the sample tested, thus, the absorbance, at 450 nm, is a measure of the concentration of CRP in the test sample. The quantity of CRP in the test sample can be interpolated from the calibration curve constructed from the calibrators, and corrected for sample dilution.

Components: 1. Diluent Concentrate: One bottle containing 50 mL of a 5X concentrated diluent running buffer.
2. Wash Solution Concentrate: One bottle containing 50 mL of a 20X concentrated wash solution.
3. Enzyme-Antibody Conjugate Concentrate: One vial containing 150 µL of a 100X concentrated affinity-purified anti-dog CRP antibody conjugated with HRP in a stabilizing buffer.
4. TMB Substrate Solution: One vial containing 12 mL of TMB and hydrogen peroxide in citric acid buffer at pH 3.3.
5. Stop Solution: One vial containing 12 mL of 0.3 M sulfuric acid. WARNING: Avoid contact with skin.
6. Microtiter Plate: Twelve removable eight-well micro strips in well holder frame. Wells are coated with affinity-purified anti-dog CRP.
7. Dog CRP Calibrator: One vial containing a lyophilized Dog CRP Calibrator.
8. Positive Control: One vial containing 50 µL of serum with 0.1% sodium azide. See the Control Certificate for the concentration.

Material not included: Precision pipettes (2 µL to 200 µL) for making and dispensing solutions
Test tubes
Microplate washer/aspirator
Distilled or de-ionized H2O Microplate reader
Assorted glassware for the preparation of reagents and buffer solutions
Timer

Application Details

Plate: Pre-coated

Reagent Preparation:

1. Diluent Concentrate: The Diluent solution supplied is a 5X concentrate and must be diluted 1:5 with distilled or de-ionized water.
   2. Wash Solution Concentrate: The Wash Solution supplied is a 20X concentrate and must be diluted 1:20 with distilled or de-ionized water. Crystal formation in the concentrate is not uncommon when storage temperatures are low. Warming of the concentrate to 30-35°C before dilution can dissolve crystals.
   3. Enzyme-Antibody Conjugate Concentrate: Calculate the required amount of working conjugate solution for each microtiter plate test strip by adding 10 µL Enzyme-Antibody Conjugate to 990 µL of 1X Diluent for each test strip to be used for testing. Mix uniformly, but gently. Avoid foaming.
   4. TMB Substrate Solution: Ready to use as supplied.
   5. Stop Solution: Ready to use as supplied.
   6. Microtiter Plate: Ready to use as supplied. Unseal Microtiter Pouch and remove plate from pouch. Remove all strips and wells that will not be used in the assay and place back in pouch and re-seal along with desiccant.
   7. Dog CRP Calibrator: Add 1.0 mL of distilled or de-ionized water to the lyophilized Dog CRP Calibrator and mix gently until dissolved. The calibrator is now at a concentration of 460.8 ng/mL (the reconstituted calibrator should be aliquoted and frozen if future use is intended). Dog CRP Calibrators need to be prepared immediately prior to use. Mix well between each step. Avoid foaming.
   8. Positive Control: The concentration and recommended dilution are provided on the Control Certificate. Before use, briefly centrifuge the Positive Control to allow all of the liquid to collect in the bottom of the vial.

Sample Preparation:

Blood should be collected by venipuncture and the serum separated from the cells, after clot formation, by centrifugation. For plasma samples, blood should be collected into a container with an anticoagulant and then centrifuged. Care should be taken to minimize hemolysis, excessive hemolysis can impact your results. Assay immediately or aliquot and store samples at -20°C. Avoid repeated freezing/thawing. For any sample that might contain pathogens, care must be taken to prevent contact with open wounds. No additives or preservatives are necessary to maintain the integrity of the specimen. Avoid azide contamination.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: 1. Complete Kit: The expiration date for the kit is stated on the outer label. The recommended storage temperature is 4°C. See long term storage recommendations for the Dog CRP Calibrator and Positive Control below.
2. Diluent: The 5X Diluent Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions should be stored at 4°C.
3. Wash Solution: The 20X Wash Solution Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions can be stored at room temperature (RT, 16-25°C) or at 4°C.
4. Enzyme-Antibody Conjugate: Undiluted anti-CRP-HRP conjugate should be stored at 4°C and diluted immediately prior to use. The working conjugate solution is stable for up to 1 hour when stored in the dark.
5. TMB Substrate Solution: The TMB Substrate Solution should be stored at 4°C and is stable until the expiration date.
6. Stop Solution: The Stop Solution should be stored at 4°C and is stable until the expiration date.
7. Microtiter Plate: Anti-dog CRP coated wells are stable until the expiration date and should be stored at 4°C in the sealed foil pouch with a desiccant pack.
8. Dog CRP Calibrator: The lyophilized Dog CRP Calibrator should be stored at 4°C or frozen until reconstituted. The reconstituted calibrator should be aliquoted and stored frozen (avoid multiple freeze-thaw cycles). The working calibrator solutions should be prepared immediately prior to use and are stable for up to 8 hours.
9. Positive Control: For storage longer than 7 days, keep frozen until the expiration date. Storage less than 7 days can be at 4°C. Avoid multiple freeze/thaw cycles.

Expiry Date: The expiry date is stated on the label.

Target Details for CRP

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

Alternative Name: HsCRP (CRP Products)

Synonyms: 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

Background: Acute phase proteins are plasma proteins, which increase in concentration following infection, inflammation or trauma. The first acute phase protein to be recognized was discovered in humans by Tillet and Frances in 1930. This CRP is so named because it is able to effect precipitation of somatic C-polysaccharide of Streptococcus pneumonia. CRP is an alpha globulin with a mass of 110,000 to 140,000 daltons, and composed of five identical subunits, which are non- covalently assembled as a cyclic pentamer. It is synthesized in the liver and, in humans, is normally present as a trace constituent of serum at a level less than 0.3 mg/dL. The levels in serum rise quickly following acute tissue damage and fall very rapidly once the stimulus is removed. It has been proposed that the function of CRP is to aid in complement activation, influence phagocytic cell function, and augment cell-mediated cytotoxicity. Investigations over the past few years have shown that quantification of CRP in plasma or serum can provide valuable information in the detection, prognosis, and monitoring of disease not only in humans, but in companion animals and farm herds as well.

Pathways: Carbohydrate Homeostasis