Strong Sensitivity Human ELISA Kit HRH4 Immunoassays Test Kit
Customized With 2 Hours Assay Time
This elisa test kit is an Enzyme-Linked Immunosorbent Assay
(ELISA). The plate has been pre-coated with Human HRH4 antibody.
HRH4 present in the sample is added and binds to antibodies coated
on the wells. And then biotinylated Human HRH4 Antibody is added
and binds to HRH4 in the sample. Then Streptavidin-HRP is added and
binds to the Biotinylated HRH4 antibody. After incubation unbound
Streptavidin-HRP is washed away during a washing step. Substrate
solution is then added and color develops in proportion to the
amount of Human HRH4. The reaction is terminated by addition of
acidic stop solution and absorbance is measured at 450 nm.
Storage: Store the reagents at 2-8°C. For over 6-month storage refer to
the expiration date keep it at -20°C. Avoid repeated thaw cycles.
If individual reagents are opened it is recommended that the kit be
used within 1 month.
*This product is for research use only, not for use in diagnosis
procedures. It’s highly recommend to read this instruction entirely
Intra-Assay Precision (Precision within an assay) Three samples of
known concentration were tested on one plate to assess intra-assay
Inter-Assay Precision (Precision between assays) Three samples of
known concentration were tested in separate assays to assess
CV(%) = SD/mean x 100
- Prior to use, the elisa test kit and sample should be warmed
naturally to room temperature 30 minutes.
- This instruction must be strictly followed in the experiment.
- Once the desired number of strips has been removed, immediately
reseal the bag to protect the remain from deterioration. Cover all
reagents when not in use.
- Make sure pipetting order and rate of addition from well-to-well
when pipetting reagents.
- Pipette tips and plate sealer in hand should be clean and
disposable to avoid cross-contamination.
- Avoid using the reagents from different batches together.
- Substrate solution B is sensitive to light, don’t expose substrate
solution B to light for a long time.
- Stop solution contains acid. Please wear eye, hand and skin
protection when using this material. Avoid contact of skin or
mucous membranes with kit reagent.
- The elisa test kit should not be used beyond the expiration date.
Serum Allow serum to clot for 10-20 minutes at room temperature.
Centrifuge at 2000-3000 RPM for 20 minutes.
Plasma Collect plasma using EDTA or heparin as an anticoagulant.
Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C
within 30 minutes of collection.
Urine Collect by sterile tube. Centrifuge at 2000-3000 RPM for
approximately 20 minutes. When collecting pleuroperitoneal fluid
and cerebrospinal fluid, please follow the procedures
Cell Culture Supernatant Collect by sterile tubes when examining secrete components.
Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect
the supernatants carefully. When examining the components within
the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the
cell concentration of approximately 1 million/ml. Damage cells
through repeated freeze-thaw cycles to let out the inside
components. Centrifuge at 2000-3000 RPM for approximately 20
Tissue and other body fluids Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly
and weigh before homogenization. Mince tissues and homogenize them
in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or
freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20
- Sample concentrations should be predicted before being used in the
assay. If the sample concentration is not within the range of the
standard curve, users must contact us to determine the optimal sample for their particular experiments.
- Samples to be used within 5 days should be stored at 2-8°C. Samples
should be aliquoted or must be stored at -20°C within 1 month or
-80°C within 6 months. Avoid repeated freeze thaw cycles.
- Samples should be brought to room temperature before starting the
- Centrifuge to collect sample before use.
- Samples containing NaN3 can’t be tested as it inhibits the activity
of Horse Radish Peroxidase (HRP).
- Collect the supernatants carefully. When sediments occurred during
storage, centrifugation should be performed again.
- Hemolysis can greatly impact the validity of test results. Take
care to minimize hemolysis.
*Sample can't be diluted with this kit. Owing to the the material we
use to prepare the kit, the sample matrix interference may falsely
depress the specificity and accuracy of the assay.
1. Prepare all reagents, samples and standards.
2. Add sample and ELISA reagent into each well. Incubate for 1 hour
3. Wash the plate 5 times.
4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.
5. Add stop solution and color develops.
6. Read the OD value within 10 minutes.
Calculation of Result
Construct a standard curve by plotting the average OD for each
standard on the vertical (Y) axis against the concentration on the
horizontal (X) axis and draw a best fit curve through the points on
the graph. These calculations can be best performed with
computer-based curve-fitting software and the best fit line can be
determined by regression analysis.