Bovine ABAT ELISA KIT

Starting at: $796.00

  • Model: ELI-13169b
  • 20 Units in Stock
Ask a question


Please Choose:


Add to Cart:

Bovine ABAT ELISA Kit



Packing.96Tests            Cat No. ELI-13169b

【INTENDED USE】
Bovine ABAT ELISA Kit is for the quantitative detection of Bovine 4-Aminobutyrate Aminotransferase Mitochondrial concentration in 
serum, plasma and other biological fluids.

This package insert must be read in its entirety before using this product.

If You Have Problems
Our expert Technical Support Staff is available to assist you in answering your questions and resolving issues to ensure complete customer satisfaction.

Please Contact Us
Nova Lifetech Inc.
Tel: (852)-81700448
Website: www.lifescience-market.com

In order to obtain higher efficiency service, please ready to supply the lot number of the kit to us (found on the outside of the box). 
 

【PRINCIPLE OF THE ASSAY】
Bovine ABAT ELISA Kit employs a two-site sandwich ELISA to quantitate ABAT in Bovine serum, plasma. An antibody specific for ABAT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ABAT present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for ABAT is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of ABAT bound in the initial step. The color development is stopped and the intensity of the color is measured.

 

【DETECTION RANGE】
0.78 ng/mL - 50 ng/mL. The standard curve concentrations used for the ELISA’s were 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.12 ng/mL, 1.56 ng/mL, 0.78 ng/mL, 0 ng/mL.

 

【SENSITIVITY】
The limit of detection of Bovine ABAT defined as the analyte concentration resulting in an absorbance significantly higher than that of the dilution medium (mean plus 2 standard deviations) was determined to be 0.39 ng/mL (mean of 6 independent assays).

 

【SPECIFICITY】
This assay has high sensitivity and excellent specificity for detection of Bovine ABAT. No significant cross-reactivity or interference between Bovine ABAT and analogues was observed.
Note:
Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between Bovine ABAT and all the analogues, therefore, cross reaction may still exist.
 

【SAMPLE COLLECTION AND STORAGE】
    Serum Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2-8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
    Plasma Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2-8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
    Other biological fluids Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at
-20°C or -80°C. Avoid repeated freeze/thaw cycles.

Note:
1.    Samples to be used within 5 days may be stored at 2-8°C, otherwise samples must be stored at -20°C (≤ 1 month) or -80°C (≤ 2 months) to avoid loss of bioactivity and contamination.
Sample hemolysis will influence the result, so hemolytic specimen cannot
be detected.
2.    When performing the assay, bring samples to room temperature.

 

【SAMPLE PREPARATION】
Bovine serum or plasma samples require no dilution before test. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments.

 

【REAGENT PREPARATION】
Bring all reagents to room temperature before use.
Wash Buffer (1 x) - If crystals have formed in the concentrate, warm up  to  room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
Biotin-Conjugate (1 x) - Centrifuge the vial before opening.
Biotin-Conjugate requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-Conjugate (100 x) + 990 μL of Biotin-Conjugate Diluent.
Streptavidin-HRP (1 x) - Centrifuge the vial before opening.
Streptavidin-HRP requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Streptavidin-HRP (100 x) + 990 μL of Streptavidin-HRP Diluent.
ABAT Standard - Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1 mL of Sample Diluent. Swirl or mix gently to insure complete and homogeneous solubilization (concentration of reconstituted standard = 50 ng/mL). The standard has to be used immediately after reconstitution and cannot be stored.
Use Eppendorf Tubes - Pipette 250 μL of the Sample Diluent into each tube. Use the stock solution to produce a dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted standard serves as the high standard (50ng/mL). The Sample Diluent serves as the zero standard (0 ng/mL).
 

【ASSAY PROCEDURE】
Bring all reagents and samples to room temperature before use. It is recommended that all samples, controls, and standards be assayed in duplicate.
1.    Prepare all reagents, working standards, and samples as directed in the previous sections.
2.    Refer to the Assay Layout Sheet to determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 2-8°C.
3.    Add 100 µL of standard and sample per well. Cover with the adhesive films provided. Incubate for 2 hours at 37°C. A plate layout is provided to record standards and samples assayed.
4.    Aspirate each well and wash, repeating the process for a total of three washes. Wash by filling each well with Wash Buffer (250 µL) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
5.    Add 100 µL of Biotin-Conjugate (1 x) to each well. Cover with the adhesive films. Incubate for 1 hour at 37°C. (Biotin-Conjugate (1 x) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)
6.    Aspirate each well and wash, repeating the process for a total of three washes. Wash by filling each well with Wash Buffer (250 µL) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
7.    Add 100 µL of Streptavidin-HRP (1 x) to each well. Cover the microtiter plate with the adhesive films. Incubate for 1 hour at 37°C.
Aspirate each well and wash, repeating the process for a total of five washes. Wash by filling each well with Wash Buffer (250 µL) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
8.    Add 100 µL of Substrate Solution to each well. Incubate for 15-20 minutes at 37°C. Keeping the plate away from drafts and other temperature fluctuations in the dark. Avoid placing the plate in direct light.
9.    Add 50 μL of Stop Solution to each well. When the first four wells containing the highest concentration of standards develop obvious blue color. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
Determine the optical density of each well within 5 minutes, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. Subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

*Samples may require dilution. See Sample Preparation section.

 

Caution:
Product is for research use only!

No customer comments for the moment.

Add A Comment

Related Products

Cart  

No products

Total $0.00

Prices don't include postage.

Cart