Dog rabies virus antibody ELISA kit
Catalog No. LSY-30012
- Principle
This kit use enzyme-linked immunoassay (ELISA) to detect specific antibody against Canine rabies virus, for monitoring antibody after dog immune.
- Reagents and contents
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Code
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Item
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Spec.
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Code
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Item
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Spec.
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1
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RBV-Ag Coated plates
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96 wells
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8
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Stop reaction solution (No.6 solution)
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1 bottle
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2
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Enzyme Conjugate
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1 bottle
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9
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Cut-off control
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1 bottle
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3
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HRPO Conjugate
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1 bottle
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10
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Negative control
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1 bottle
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4
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PBST powder
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2 bags
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11
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Adhesive Foil
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2 pieces
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5
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Substrate A (No.3 solution)
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1 bottle
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12
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Sealed bag
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1 piece
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6
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Substrate B(No.4 solution)
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1 bottle
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13
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Instruction sheet
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1 piece
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7
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Sample diluent (No.5 solution)
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1 bottle
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3. Material required not provided
1) Microplate Reader with 450nm and 630nm
2) 37 ℃thermostatic device
3) Micropipettes, adjustable.
4. Sample requirement
1) To get the best test result, avoid using sample with severe hemolysis, precipitate, contaminated by bacteria or protein suspension.
2) The serum sample storage time should be less than 1 week at 2-8 ℃, if for long term, it should be kept at -20℃ or lower.
5. Test procedure
1. Conjugate working solution preparation: add one Enzyme Conjugate into one HRPO Conjugate, mix thoroughly;
2. Washing Solution preparation: Dissolve each bag PBST powder with 500 ml distilled water,
3. Adding samples and controls: Add two drops Sample diluent into each well, then add 5 μl serum sample into each well separately, mix evenly. Set 2 wells for negative control, 2 wells for Cut-off control, 1 well for blank control. (Directly add negative control and Cut-off control to its corresponding wells, 100ul/well, blank control well only drop 2 drops sample diluent). Incubate in dark at 37 ℃ for 20 minutes, then discard the liquid of the well, add 1 drop washing solution into each well and fill with distilled water promptly, incubate for 30 seconds, then discard the liquid, continue perform another 4 washing cycles as above, then discard the liquid, flap to dry with the absorbent paper.
4. Adding conjugate working solution: Dispense 2 drops into all wells (except blank well) and incubate at 37℃ for 20 minutes. Discard the contents of the wells and wash 5 times as described in step 3.
5. Adding substrates: Add 1 drop Substrate A (No.3 solution) and 1 drop Substrate B (No.4 solution) into all well, incubate at 37℃ for 10 minutes.
6. Add one drop Stop reaction solution (No.6 solution) into all wells to stop reaction. Set zero at blank control, read the OD value at ELISA reader 450nm (630nm as a reference).
6. Reference
Validation judge
Blank control OD Value < 0.05 (If more than 0.05, the test is invalid)
Negative control OD Value ≤ 0.15 (If more than 0.15, the test is invalid
Cut-off control OD value ≥ 0.20 (If less than 0.20, the test is invalid)
7. Results interpretation
Sample OD value ≥ cut-off control: positive, it means the antibody level≥0.5IU/ ml.
Sample OD value < cut-off control: negative, it means the antibody level <0.5IU/ml.
8. Limitation
This kit is only suitable for dog serum
9. Notes
1) RBV-Ag coated plates should be sealed and moisture-proof. MicroWell plate removed from the refrigerated environment should be balanced moisture to dry at room temperature, then can be opened. Put back unused MicroWell plate into dry foil bag and sealed at 4 ℃.
2) Before dropping reagent, gently shake dropping bottle and mix even the reagent, discard the first drop; Make sure the dropping bottle is vertical when dropping.
3) Screw down the cap after use, don't mix caps between different bottles and don't mix components between different kits
4) When incubate at 37℃, suggested to use water bath. If there is no water bath, put wet box with adhesive foil in 37℃ incubator.
5) The test kit and Any samples should be regarded as the source of infection to properly handle. Stop solution is corrosive, be careful to use.
10. Storage and expire date
- Store at 2~8℃ in dark, no frozen, expiry date: 12 months.
- After open the RBV-Ag coated plates, store at 2~8℃ in dark, the period of use is 15 days.
Shenzhen Lvshiyuan Biotechnology Co., Ltd
D Building, National Biological Industrial Park of Marinelife, No.2 Binhai Road, Dapeng, Shenzhen, 518120 China
Tel. 86-755-28438788
Fax 86-755-28938800
www.lsybt.com
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