Shanghai Korain Biotech Co., Ltd |
|
Canine VACHT Immunosorbent Assay Kit Vesicular Acetylcholine
Transporter ELISA Kit For Research
Cat.No E0289Ca
Intended Use
This sandwich kit is for the accurate quantitative detection of
Canine Vesicular Acetylcholine Transporter (also known as VACHT) in
serum, plasma, cell culture supernates, cell lysates, tissue
homogenates.
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 ELISA Assay Kit is an Enzyme-Linked Immunosorbent Assay
(ELISA). The plate has been pre-coated with Canine VACHT antibody.
VACHT present in the sample is added and binds to antibodies coated
on the wells. And then biotinylated Canine VACHT Antibody is added
and binds to VACHT in the sample. Then Streptavidin-HRP is added
and binds to the Biotinylated VACHT 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 Canine VACHT. The reaction is terminated by
addition of acidic stop solution and absorbance is measured at 450
nm.
Standard Curve Range: 0.05ng/ml - 20ng/ml
Sensitivity: 0.023ng/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
Components | Quantity |
Standard Solution (24ng/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 (25x) | 20ml x1 |
Biotinylated Canine VACHT Antibody | 1ml x1 |
User Instruction | 1 |
Plate Sealer | 2 pics |
Zipper bag | 1 pic |
Material Required But Not Supplied
Precautions
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-VACHT
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.
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.