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SP Sandwich Canine ELISA Kits High Specificity Canine Substance P With Oem Service

Categories Canine ELISA Kits
Brand Name: BT Lab
Model Number: Cat.No E0288Ca
Certification: CE, ISO9001:2005, 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
Shipping: DHL/FedEX
Bulk Order: Yes
Quality: CE, ISO
Target Protein: Substance P
Delivery: Within 48 hours
Test Method: Sandwich
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SP Sandwich Canine ELISA Kits High Specificity Canine Substance P With Oem Service

SP Sandwich ELISA Kit High Specificity Canine Substance P ELISA Test Kit With Oem Service


Cat.No E0288Ca

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 and 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.


*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.

Standard Curve Range: 0.2ng/ml - 70ng/ml

Sensitivity: 0.13ng/ml

Size: 96 wells

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.


Reagent Provided

ComponentsQuantity
Standard Solution (80ng/ml)0.5ml x1
Pre-coated ELISA Plate12 * 8 well strips x1
Standard Diluent3ml x1
Streptavidin-HRP6ml x1
Stop Solution6ml x1
Substrate Solution A6ml x1
Substrate Solution B6ml x1
Wash Buffer Concentrate (25x)20ml x1
Biotinylated Canine SP Antibody1ml x1
User Instruction1
Plate Sealer2 pics
Zipper bag1 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

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

CV(%) = SD/mean x 100

Intra-Assay: CV<8%

Inter-Assay: CV<10%


Intended Use

This sandwich kit is for the accurate quantitative detection of Canine Substance P (also known as SP) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.


Reagent Preparation

All reagents should be brought to room temperature before use.

Standard Reconstitute the 120μl of the standard (80ng/ml) with 120μl of standard diluent to generate a 40ng/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 (40ng/ml) 1:2 with standard diluent to produce 20ng/ml, 10ng/ml, 5ng/ml and 2.5ng/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:


40ng/mlStandard No.5120μl Original Standard + 120μl Standard Diluent
20ng/mlStandard No.4120μl Standard No.5 + 120μl Standard Diluent
10ng/mlStandard No.3120μl Standard No.4 + 120μl Standard Diluent
5ng/mlStandard No.2120μl Standard No.3 + 120μl Standard Diluent
2.5ng/mlStandard No.1120μl Standard No.2 + 120μl Standard Diluent

Standard ConcentrationStandard No.5Standard No.4Standard No.3Standard No.2Standard No.1
80ng/ml40ng/ml20ng/ml10ng/ml5ng/ml2.5ng/ml

Wash Buffer Dilute 20ml of Wash Buffer Concentrate 25x into deionized or distilled water to yield 500 ml of 1x Wash Buffer. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved.


Summary

1. Prepare all reagents, samples and standards.

2. Add sample and ELISA reagent into each well. Incubate for 1 hour at 37°C.

3. Wash the plate 5 times.

4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.

5. Add stop solution and color develops.

6. Read the OD value within 10 minutes.


Calculation of Result

Construct a standard curve by plotting the average OD for each standard on the vertical (Y) axis against the concentration on the horizontal (X) axis and draw a best fit curve through the points on the graph. These calculations can be best performed with computer-based curve-fitting software and the best fit line can be determined by regression analysis.

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