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Shanghai Korain Biotech Co Ltd
8 Years
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| Brand Name : | BT Lab |
| Model Number : | Cat.No E0166Ra |
| Certification : | CE, ISO9001:2005, MSDS |
| Price : | Negotiation |
| Supply Ability : | Western Union, T/T |
| Delivery Time : | 1-3 business days, bulk order within one week |
96 Wells 48 Wells Rat IFABP ELISA Kit Strong Sensitivity For
Accurate Quantitative Detection
Cat.No E0166Ra
Standard Curve Range: 1ng/ml - 400ng/ml
Sensitivity: 0.52ng/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.
Intended Use
This sandwich kit is for the accurate quantitative detection of Rat
Intestinal Fatty Acid Binding Protein (also known as IFABP) in
serum, plasma, cell culture supernates, cell lysates, tissue
homogenates.
Assay Principle
This kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate
has been pre-coated with Rat IFABP antibody. IFABP present in the
sample is added and binds to antibodies coated on the wells. And
then biotinylated Rat IFABP Antibody is added and binds to IFABP in
the sample. Then Streptavidin-HRP is added and binds to the
Biotinylated IFABP 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 IFABP. The reaction is terminated by addition of
acidic stop solution and absorbance is measured at 450 nm.
Reagent Provided
| Components | Quantity |
| Standard Solution (480ng/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 Rat IFABP Antibody | 1ml x1 |
| User Instruction | 1 |
| Plate Sealer | 2 pics |
| Zipper bag | 1 pic |
Material Required But Not Supplied
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (480ng/ml) with 120μl of
standard diluent to generate a 240ng/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 (240ng/ml) 1:2 with standard
diluent to produce 120ng/ml, 60ng/ml, 30ng/ml and 15ng/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:
| 240ng/ml | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 120ng/ml | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 60ng/ml | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 30ng/ml | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 15ng/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 |
| 480ng/ml | 240ng/ml | 120ng/ml | 60ng/ml | 30ng/ml | 15ng/ml |
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.
Referances
"Chronic leptin administration decreases fatty acid uptake and
fatty acid transporters in rat skeletal muscle."
Steinberg G.R., Dyck D.J., Calles-Escandon J., Tandon N.N., Luiken
J.J., Glatz J.F., Bonen A.
J. Biol. Chem. 277:8854-8860(2002)
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