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Avian Leukosis Antigen ELISA Kit (ADES0032)

SKU:
ADES0032
Product Type:
ELISA Kit
Size:
96 Assays
Assay Type:
Sandwich ELISA
Reactivity:
Poultry
€599
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Description

system_update_altDatasheetsystem_update_altMSDS
Product Name: Avian Leukosis Antigen ELISA Kit
SKU: ADES0032
Target: ALV-P27-Ag
Size: 96T
Assay type: Sandwich-ELISA
Assay time: 1.10h
Sample Type: Cloacal orifice, Yolk
Reactivity: Poultry
Detection Range: 450/630nm
Result Type: Qualitative; Sensitivity 98.5%; Specificity 99.5%
Kit component:
Component Quantity (96 Assays)
Micro ELISA Plate (Dismountable) 96 Wells
HRP Conjugate 25 mL
20×Concentrated Wash Buffer 25 mL
Substrate Reagent A 15 mL
Substrate Reagent B 15 mL
Stop Solution 15 mL
Positive Control 2 mL
Negative Control 2 mL
Plate Sealer 3 Pieces
Sealed Bag 1 Piece
Product Description 1 copy

This kit is comprised by HRP conjugate, other reagents and ELISA Microtiter plate pre-coated with Avian Leukosis (ALV) P27 antibodies. Apply the principle of enzyme-linked immunoassay (ELISA) to detect ALV-Ag in cloacal orifice, yolk of poultry. During the experiment, add control and samples into the ELISA Microtiter plate, ALV-Ag will be bound with the antibodies on the ELISA Microtiter plate. Then wash the plate to remove unbound components, horseradish peroxidase (HRP) conjugate is added to each ELISA Microtiter plate well. The unbound HRP Conjugate will be removed by washing and substrate reagent is added for color development. At last, end the reaction by adding Stop Solution to produce a yellow product. There is a positive correlation between the OD value of samples and the concentration of ALV-Ag. Measure the absorbance value of each well by using a microplate reader with 450 nm (630 nm) wavelength, then we can judge whether ALV antibody exist in the sample.

Proper solution and sample preparation are essential to ensure accurate and reliable results. Below are the steps for preparing solutions and samples for analysis.

Preparation Type Protocol
Solution Preparation
  • Wash Buffer: Adjust the 25× Concentrated Wash Buffer to room temperature before use. Shake well to dissolve any salt crystals that may appear. Dilute with deionized water at a 1:24 ratio (e.g., 1 mL of 25× Concentrated Wash Buffer + 24 mL deionized water).
  • 0.01M PBS Solution (pH=7.4): Dissolve 8.0 g of NaCl, 0.2 g of KCl, 1.14 g of Na2HPO4, and 0.24 g of KH2PO4 in 1000 mL of deionized water. This solution can be stored at room temperature for long periods.
Sample Preparation - Cloacal Orifice
  1. Insert the cotton swab into the cloacal orifice and rotate repeatedly on the inner wall to collect the sample (adequate sample collection affects detection results).
  2. Immediately insert the cotton swab into a tube containing 1 mL of 0.01 M PBS (pH=7.4). Stir the swab until the sample is fully dissolved into the diluent. Discard the swab after wiping it against the tube wall.
  3. Freeze the sample tube once at a temperature below -20 °C. Thaw the sample tube to room temperature before analysis.
  4. Take 100 μL of the sample solution for analysis.
Sample Preparation - Yolk Take 100 μL of the yolk sample for analysis.

Follow the steps below for conducting the assay. Ensure all procedures are performed carefully to maintain accuracy and reliability.

Step Protocol
1. Number Number the sample and control wells in order (for multiple wells) and record the positions of control and sample wells. Set 2 wells each for negative and positive controls. Test samples in duplicate.
2. Add Sample Add 100 μL of positive control to positive control wells and 100 μL of negative control to negative control wells. Add 100 μL of sample to sample wells.
3. Incubate Cover the plate with a plate sealer and mix thoroughly. Incubate at 37°C for 30 minutes in the dark.
4. Wash Remove the liquid from each well. Immediately add 260 μL of Wash Buffer to each well and wash. Repeat the wash procedure 5 times with 30-second intervals between washes. After washing, invert the plate and tap it against thick, clean absorbent paper. If bubbles are present, use clean tips to remove them.
5. HRP Conjugate Add 100 μL of HRP conjugate to each well. Cover the plate with a plate sealer and incubate at 37°C for 30 minutes in the dark.
6. Wash Repeat Step 4 for washing.
7. Color Development Add 50 μL of Substrate Reagent A and 50 μL of Substrate Reagent B to each well. Mix thoroughly, cover the plate with a sealer, and incubate at 37°C for 10 minutes in the dark.
8. Stop Reaction Add 50 μL of Stop Solution to each well and mix thoroughly.
9. OD Measurement Measure the absorbance value (A-value) of each well using a Microplate Reader at a wavelength of 450 nm (use 630 nm as the reference wavelength). Note: Read results within 5 minutes.

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