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Rabies Virus Antibody ELISA Kit for Pet 96 Wells/Kit GMP ISO9001
1. Introduction
The Rabies Virus (RBV) antibody ELISA kit is used to test rabies
virus antibody in serum of dogs, cats etc., used to assess the
status of rabies vaccination.
This kit use block ELISA method, rabies antigen is pre-coated on
enzyme micro-well strips. When testing, add diluted serum sample,
after incubation, if there is rabies virus specific antibody, it
will combine with the pre-coated antigen, discard the uncombined
antibody and other components with washing; then add enzyme labled
anti-rabies virus monoclonal antibody, antibody in sample block the
combination of monoclonal antibody and pre-coated antigen; discard
the uncombined enzyme conjugate with washing; Add TMB substrate in
micro-wells, the blue signal by Enzyme catalysis is in inverse
proportion of antibody content in sample, use ELISA reader at 450nm
wavelenth to measure the absorbance A value in reaction wells after
adding stop solution to stop the reaction.
2. Reagents and contents
Code | Item | Spec. | Code | Item | Spec. |
1 | RBV-AgCoatedplates | 1/2 plate of 96 wells | 6 | Stop solution | 11 ml |
2 | EnzymeConjugate | 11/22 ml | 7 | Negative control | 1/2ml |
3 | 10X Concentrated washing buffer | 100 ml | 8 | Positive control | 0.5/1 ml |
4 | Substrate | 11/22 ml | 9 | Adhesive Foil | 2/4 pieces |
5 | Sample diluent | 100 ml | 10 | Instruction sheet | 1 piece |
3. Materialrequired notprovided
1) Micropipette: 10ul-100ul, 100ul-1000ul.
2) Disposable pipette tips.
3) Graduate: 500ml.
4) Microplate Reader: 96 wells.
5) Distilled water or deionized water.
6) Microplate Washer
4. Sample preparation
Take animal whole blood, get serum by using regular method, the
serum should bright and no hemolysis
5. Washing buffer preparation
Return 10X Concentrated washing buffer into room temperature before
use, if there is salt crystals, shake to make it dissolved, then
dilute it at 10 times with distilled water or deionized water. The
diluted washing buffer can store at 4℃for about 1 week.
6. Notes
1) Return all reagents into room temperature before use, shake it
evenly before use, and store back to 2-8℃after usage.
2) Do not mix use reagents from different kits and different lot
no., prevent the reagents been polluted when using.
3) Substrate A and stop solution may have irritation to skin and
eyes, be careful to use.
4) Do not expose Substrate to strong light and avoid contact with
the oxidant.
5) RBV-Ag coated plates should be sealed and moisture-proof. Put
back unused MicroWell plate into dry foil bag and sealed at 2~8 ℃.
6) All wastes should be treated well to avoid pullution before
discarding.
7) Strict compliance with the operating instructions can get the
best results. Pipetting operation, timing, and washing of the whole
process must be precise.
8) RBV-Ag coated plates is disposable, do not repeat use.
7. Test procedure
1) For every test, set 1 well for positive control and 2 wells for
negative control, negative control and positive control do not need
to dilute, directly add to its corresponding wells, 100ul/well;
2) Add sample diluent into reaction wells, 80ul/well, then add
serum, 20ul/well, blow and beat it to even. (Do not mix use the
tips);
3) Cover it with Adhesive Foil,incubateat 37 ℃ for 30 minutes;
4) Open the adhesive foil, discard the liquid of the well, add
diluted washing buffer to each well, 250ul/well, discard the
liquid, repeat the above step for 5 times, at last flap to dry with
the absorbent paper;
5) Adding Enzyme Conjugate,100ul/well, cover it with Adhesive
Foil,incubateat 37 ℃ for30 minutes;
6) Open the adhesive foil, discard the liquid of the well, washing
for 5 times as step 4), remember at last flap to dry with the
absorbent paper;
7) Add substrate, 100ul/well, mix it evenly then cover it with
Adhesive Foil,incubateat 37 ℃in darkfor15 minutes;
8) Add stop solution, 50ul/well to stop the reaction, measure the
result in 10 minutes.
8. Results judgment
Read the OD value at 450nm (630nm as reference).
For the assay to be valid:
OD value of negative control(N) > 0.5, meanwhile positive value
(P) blocking rate > 60%
Calculate method:
PI(blocking rate)= 1- (Sample OD value/ Negative control OD average
value)
Resultsinterpretation
PI(blocking rate)> 60%: Positive
PI(blocking rate)≤60%: Negative
(Note: when PI=60%, it means antibody titer 0.5 IU/ml )
9. Storage and expire date
Store at 2~8℃ in dark, no frozen, expiry date: 12 months.
Q1: How long will it take for you to ship?
A1: Usually ships within 7 working days.
Q2: Do you support OEM/ODM?
A2: can be supported. We can customize according to your specific
needs and specific quantities.
Q3: How is your factory doing in terms of quality control?
A3: We have ISO9001 certification and ISO13485 certification, so
far all the production process, we have standard rules, we comply
with the relevant behavior and laws of the government.
Q4: Is after-sales service guaranteed?
A4: We provide professional online technical after-sales service.
If the product fails during the experiment, we can provide
one-to-one guidance through telephone, video and other forms.
Q5: What is your minimum order quantity?
A5: 1Kit.
Q6: What is the shipping method?
A6: It can be shipped by express (FEDEX, UPS, DHL, EMS, etc.) or by
air and ground.Please confirm with us before placing an order.