Rat BFGF Sandwich ELISA Kit Basic Fibroblast Growth Factor ELISA Kit With 2 Hours Assay Time

Sandwich Type Rat BFGF ELISA Test Kit Basic Fibroblast Growth Factor ELISA Kit With 2 Hours Assay Time

Cat.No E0341Ra

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

Sensitivity: 1.16ng/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. It’s highly recommend to read this instruction entirely before use.

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.

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 Rat Basic Fibroblast Growth Factor (also known as BFGF) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.

Assay Principle

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

Precautions

• Prior to use, the sandwich 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 sandwich kit should not be used beyond the expiration date.

Reagent Provided

Reagent Preparation

All reagents should be brought to room temperature before use.

Standard Reconstitute the 120μl of the standard (640ng/L) with 120μl of standard diluent to generate a 320ng/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 (320ng/L) 1:2 with standard diluent to produce 160ng/L, 80ng/L, 40ng/L and 20ng/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:

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.