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Beta Defensin 2 Mouse Elisa Kit High Sensitivity 96 Wells With Oem Service

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Beta Defensin 2 Mouse Elisa Kit High Sensitivity 96 Wells With Oem Service

Beta Defensin 2 Mouse Elisa Kit High Sensitivity 96 Wells With Oem Service
Beta Defensin 2 Mouse Elisa Kit High Sensitivity 96 Wells With Oem Service

Large Image :  Beta Defensin 2 Mouse Elisa Kit High Sensitivity 96 Wells With Oem Service

Product Details:

Place of Origin: Shanghai, China
Brand Name: BT Lab
Certification: CE, ISO9001:2005, MSDS
Model Number: Cat.No E1738Mo

Payment & Shipping Terms:

Minimum Order Quantity: Negotiation
Price: Negotiation
Packaging Details: Wrapped with ice pack and styrofoam package
Delivery Time: 1-3 business days, bulk order within one week
Payment Terms: Western Union, T/T
Supply Ability: In Stock
Detailed Product Description
OEM: Acceptable Custom: Available
Lead Time: Within 48 Hours Sample: Serum,plasma,urine,tissue,cell Culture Supernatant
Quality: CE, ISO Shipping: DHL/FedEX
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96 Wells Customized Mouse Beta Defensin 2 ELISA Kit High Sensitivity

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Cat.No E1738Mo

Standard Curve Range: 10ng/L - 2000ng/L

Sensitivity: 5.06ng/L

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. Its highly recommend to read this instruction entirely before the use.

 

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%

 

Assay Principle

This kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate has been pre-coated with Mouse DEFB2 antibody. DEFB2 present in the sample is added and binds to antibodies coated on the wells. And then biotinylated Mouse DEFB2 Antibody is added and binds to DEFB2 in the sample. Then Streptavidin-HRP is added and binds to the Biotinylated DEFB2 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 Mouse DEFB2. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.

 

Reagent Provided

Components Quantity
Standard Solution (2400ng/L) 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 Mouse DEFB2 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

 

Precautions

  • Prior to use, the kit and sample should be warmed naturally to room temperature 30 minutes.
  • This instruction must be strictly followed in the experiment.
  • Once the desired number of strips has been removed, immediately reseal the bag to protect the remain from deterioration. Cover all reagents when not in use.
  • Make sure pipetting order and rate of addition from well-to-well when pipetting reagents.
  • Pipette tips and plate sealer in hand should be clean and disposable to avoid cross-contamination.
  • Avoid using the reagents from different batches together.
  • Substrate solution B is sensitive to light, don’t expose substrate solution B to light for a long time.
  • Stop solution contains acid. Please wear eye, hand and skin protection when using this material. Avoid contact of skin or mucous membranes with kit reagent.
  • The kit should not be used beyond the expiration date.

 

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 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.

 

Reagent Preparation

All reagents should be brought to room temperature before use.Standard Reconstitute the 120μl of the standard (2400ng/L) with 120μl of standard diluent to generate a 1200ng/L 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 (1200ng/L) 1:2 with standard diluent to produce 600ng/L, 300ng/L, 150ng/L and 75ng/L solutions. Standard diluent serves as the zero standard(0 ng/L). Any remaining solution should be frozen at -20°C and used within one month. Dilution of standard solutions suggested are as follows:

 

1200ng/L Standard No.5 120μl Original Standard + 120μl Standard Diluent
600ng/L Standard No.4 120μl Standard No.5 + 120μl Standard Diluent
300ng/L Standard No.3 120μl Standard No.4 + 120μl Standard Diluent
150ng/L Standard No.2 120μl Standard No.3 + 120μl Standard Diluent
75ng/L 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
2400ng/L 1200ng/L 600ng/L 300ng/L 150ng/L 75ng/L

 

Wash Buffer Dilute 20ml of Wash Buffer Concentrate 30x 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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Shanghai Korain Biotech Co., Ltd

Contact Person: Lee

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