Porcine Foot and Mouth Disease Virus 3ABC Antibody ELISA Test Kit
192 Wells/Kit
1. Principle
Foot-and-mouth disease virus (FMDV) Non-structural protein Antibody
3ABC ELISA Test Kit is suitable for test serum of cattle, sheep,
goats and pigs, it can distinguish between immunized animals and
wild-infected animals.
This kit use block ELISA method, FMDV-3ABC antigen is pre-coated on
enzyme micro-well strips. When testing, add diluted serum sample,
after incubation, if there is FMD 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-FMD 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 | FMDV-3ABC Ag Coated plates 96 wells | 2 plates | 6 | Stop solution | 11 ml |
| 2 | Enzyme Conjugate | 22 ml | 7 | Negative control | 2 ml |
| 3 | 10XConcentrated Washing buffer | 100 ml | 8 | Positive control | 1 ml |
| 4 | Substrate | 22 ml | 9 | Adhesive Foil | 4 pieces |
| 5 | Sample diluent | 50 ml | 10 | Instruction | 1 piece |
3. Material required not provided
1) Micropipette: 10ul-100ul, 100ul-1000ul.
2) Disposable pipette tips.
3) Graduate: 500ml.
4) Microplate Reader: 96 wells with 450/630nm wavelength.
5) Distilled water or deionized water.
6) Bottle washer or 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, put the
reagents at room temperature for at least 1 hour. 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 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) FMDV-3ABC Ag coated plates should be sealed and moisture-proof.
Put back unused Micro-Well plate into dry foil bag and sealed at
2-8 ℃.
6) All wastes should be treated well to avoid pollution 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) FMDV-3ABC Ag Coated plates is disposable, do not repeat use.
7. Test procedure
1) Take the antigen coated plate(the plate can be open and used for
several times according to sample quantity each time), for every
test, set 1 well for positive control and 2 wells for negative
control, positive control and negative control do not need dilute,
take 100ul directly and add into its well;
2) Add Sample diluent to reaction wells, 80ul/well, then add serum
sample to the reaction wells, 20ul/well, blow and mix evenly(Do not
mix use tips)
3) Cover it with Adhesive Foil, incubate at 37 ℃ for 60 minutes;
4) Open the adhesive foil, discard the liquid of the well, add
diluted washing buffer to each well, 250ul/well, then discard the
liquid, repeat the above step for 4-6 times, at last flap to dry
with the absorbent paper;
5) Adding Enzyme Conjugate 100ul/well, Cover it with Adhesive Foil,
incubate at 37 ℃ for 60 minutes;
6) Open the adhesive foil, discard the liquid of the well, washing
for 4-6 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, incubate at 37 ℃ in dark for 15 minutes;
8) Add stop solution 50ul/well to stop the reaction, measure the
result in 10 minutes.
8. Results judgment
Read the OD value with ELISA Reader at 450nm (630nm as reference).
For the assay to be valid:
Negative control (N) OD value > 0.6 , meanwhile positive control
(PI) blocking rate > 50%;
Calculate method:
PI(blocking rate) = 1 - (Sample OD value/Average OD value of
Negative control)
Results interpretation
PI(blocking rate) > 50%: Positive
PI(blocking rate)≤ 50%: Negative
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: Usually by air, we will choose the best shipping method for
you, and we can also ship according to the shipping method you
require.
