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Strong Sensitivity Bovine Prolactin Sandwich Immunoassay Kit With Strong Sensitivity

Shanghai Korain Biotech Co., Ltd

Strong Sensitivity Bovine Prolactin Sandwich Immunoassay Kit With Strong Sensitivity

Brand Name : BT Lab

Model Number : Cat.No E0237Bo

Certification : CE, ISO9001:2015, MSDS

Place of Origin : Shanghai, China

MOQ : Negotiation

Price : Negotiation

Supply Ability : Western Union, T/T

Delivery Time : 1-3 business days, bulk order within one week

Packaging Details : Wrapped with ice pack and styrofoam package

Assay Principle : Sandwich

In Stock : Yes

Assay Length : 2 hours

Sensitivity : 1.15ng/ml

Standard Curve Range : 2ng/ml - 600ng/ml

Customized : Acceptable

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Strong Sensitivity Bovine Prolactin Sandwich Immunoassay Kit With Strong Sensitivity

Cat.No E0237Bo

Size: 96 wells

Intended Use

This sandwich kit is for the accurate quantitative detection of Bovine Prolactin (also known as PRL) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.

Storage: Store the reagents at 2-8°C. For over 6-month storage refer to the expiration date keep it at -20°C. Avoid repeated thaw cycles. If individual reagents are opened it is recommended that the kit be used within 1 month.

*This product is for research use only, not for use in diagnosis procedures. It’s highly recommend to read this instruction entirely before use.

Assay Principle

This ELISA Test Kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate has been pre-coated with Bovine PRL antibody. PRL present in the sample is added and binds to antibodies coated on the wells. And then biotinylated Bovine PRL Antibody is added and binds to PRL in the sample. Then Streptavidin-HRP is added and binds to the Biotinylated PRL antibody. After incubation unbound Streptavidin-HRP is washed away during a washing step. Substrate solution is then added and color develops in proportion to the amount of Bovine PRL. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.

Reagent Provided

Components Quantity
Standard Solution (800ng/ml) 0.5ml x1
Pre-coated ELISA Plate 12 * 8 well strips x1
Standard Diluent 3ml x1
Streptavidin-HRP 6ml x1
Stop Solution 6ml x1
Substrate Solution A 6ml x1
Substrate Solution B 6ml x1
Wash Buffer Concentrate (30x) 20ml x1
Biotinylated Bovine PRL Antibody 1ml x1
User Instruction 1
Plate Sealer 2 pics
Zipper bag 1 pic

Material Required But Not Supplied

  • 37°C±0.5°C incubator
  • Absorbent paper
  • Precision pipettes and disposable pipette tips
  • Clean tubes
  • Deionized or distilled water
  • Microplate reader with 450 ± 10nm wavelength filter

Specimen Collection

Serum Allow serum to clot for 10-20 minutes at room temperature. Centrifuge at 2000-3000 RPM for 20 minutes.

Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C within 30 minutes of collection.

Urine Collect by sterile tube. Centrifuge at 2000-3000 RPM for approximately 20 minutes. When collecting pleuroperitoneal fluid and cerebrospinal fluid, please follow the procedures above-mentioned.

Cell Culture Supernatant Collect by sterile tubes when examining secrete components. Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect the supernatants carefully. When examining the components within the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the cell concentration of approximately 1 million/ml. Damage cells through repeated freeze-thaw cycles to let out the inside components. Centrifuge at 2000-3000 RPM for approximately 20 minutes.

Tissue other body fluids Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly and weigh before homogenization. Mince tissues and homogenize them in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20 minutes.

Note

  • Sample concentrations should be predicted before being used in the assay. If the sample concentration is not within the range of the standard curve, users must contact us to determine the optimal sample for their particular experiments.
  • Samples to be used within 5 days should be stored at 2-8°C. Samples should be aliquoted or must be stored at -20°C within 1 month or -80°C within 6 months. Avoid repeated freeze thaw cycles.
  • Samples should be brought to room temperature before starting the assay.
  • Centrifuge to collect sample before use.
  • Samples containing NaN3 can’t be tested as it inhibits the activity of Horse Radish Peroxidase (HRP).
  • Collect the supernatants carefully. When sediments occurred during storage, centrifugation should be performed again.
  • Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.

*Sample can't be diluted with this kit. Owing to the the material we use to prepare the kit, the sample matrix interference may falsely depress the specificity and accuracy of the assay.

Reagent Preparation

All reagents should be brought to room temperature before use.

Standard Reconstitute the 120μl of the standard (800ng/ml) with 120μl of standard diluent to generate a 400ng/ml standard stock solution. Allow the standard to sit for 15 mins with gentle agitation prior to making dilutions. Prepare duplicate standard points by serially diluting the standard stock solution (400ng/ml) 1:2 with standard diluent to produce 200ng/ml, 100ng/ml, 50ng/ml and 25ng/ml solutions. Standard diluent serves as the zero standard(0 ng/ml). Any remaining solution should be frozen at -20°C and used within one month. Dilution of standard solutions suggested are as follows:

400ng/ml Standard No.5 120μl Original Standard + 120μl Standard Diluent
200ng/ml Standard No.4 120μl Standard No.5 + 120μl Standard Diluent
100ng/ml Standard No.3 120μl Standard No.4 + 120μl Standard Diluent
50ng/ml Standard No.2 120μl Standard No.3 + 120μl Standard Diluent
25ng/ml Standard No.1 120μl Standard No.2 + 120μl Standard Diluent

Standard Concentration Standard No.5 Standard No.4 Standard No.3 Standard No.2 Standard No.1
800ng/ml 400ng/ml 200ng/ml 100ng/ml 50ng/ml 25ng/ml

Wash Buffer Dilute 20ml of Wash Buffer Concentrate 30x into deionized or distilled water to yield 600 ml of 1x Wash Buffer. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved.

Assay Procedure

1. Prepare all reagents, standard solutions and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature.

2. Determine the number of strips required for the assay. Insert the strips in the frames for use. The unused strips should be stored at 2-8°C.

3. Add 50μl standard to standard well. Note: Don’t add antibody to standard well because the standard solution contains biotinylated antibody.

4. Add 40μl sample to sample wells and then add 10μl anti-PRL antibody to sample wells, then add 50μl streptavidin-HRP to sample wells and standard wells (Not blank control well). Mix well. Cover the plate with a sealer. Incubate 60 minutes at 37°C.

5. Remove the sealer and wash the plate 5 times with wash buffer. Soak wells with at least 0.35 ml wash buffer for 30 seconds to 1 minute for each wash. For automated washing, aspirate all wells and wash 5 times with wash buffer, overfilling wells with wash buffer. Blot the plate onto paper towels or other absorbent material.

6. Add 50μl substrate solution A to each well and then add 50μl substrate solution B to each well. Incubate plate covered with a new sealer for 10 minutes at 37°C in the dark.

7. Add 50μl Stop Solution to each well, the blue color will change into yellow immediately.

8. Determine the optical density (OD value) of each well immediately using a microplate reader set to 450 nm within 10 minuets after adding the stop solution.


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