Salmon COMPLEMENT 3 ELISA Kit
Packing.96Tests Cat No. ELA-E0861s
【INTENDED USE】
For the quantitative detection of Salmon Complement 3 concentration in serum, plasma and other biological fluids.
This package insert must be read in its entirety before using this product.
If You Have Problems
Our expert Technical Support Staff is available to assist you in answering your questions and resolving issues to ensure complete customer satisfaction.
Please Contact Us
Nova Lifetech Inc.
Tel: (852)-81700345
Website: www.lifescience-market.com
In order to obtain higher efficiency service, please ready to supply the lot number of the kit to us (found on the outside of the box).
【REAGENTS AND MATERIALS PROVIDED】
Reagents
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Quantity
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Reagents
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Quantity
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Assay plate (96 Wells)
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1
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Instruction manual
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1
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Standard (lyophilized)
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2
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Sample Diluent
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1 x 20 mL
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Biotin-Conjugate (concentrate 100 x)
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1 x 120μL
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Biotin-Conjugate Diluent
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1 x 20 mL
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Streptavidin-HRP (concentrate 100 x)
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1 x 120μL
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Streptavidin-HRP Diluent
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1 x 20 mL
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Wash Buffer (concentrate 25 x)
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1 x 20 mL
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Substrate Solution
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1 x 12 mL
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Stop Solution
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1 x 10 mL
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Adhesive Films
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4
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【MATERIALS REQUIRED BUT NOT SUPPLIED】
※ Microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
※ Precision single or multi-channel pipettes and disposable tips.
※ Deionized or distilled water.
※ Eppendorf Tubes for serial dilution samples.
※ Container for Wash Solution.
※ Absorbent paper for blotting the microtiter plate.
【STORAGE】
Unopened kit
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Store at 2 - 8°C. Do not use past kit in expiration date.
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Opened/ Reconstituted Reagents
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Coated assay plate
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May be stored for up to 1 month at 2 - 8°C. Return unused wells to the foil pouch containing the desiccant pack, reseal along entire edge of zip-seal, and avoid the damp.
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Standard
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May be stored for up to 1 month at 2 - 8°C. If not for recent use, better keep it store at -20°C.
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Biotin-Conjugate
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Streptavidin-HRP
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Sample Diluent
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May be stored for up to 1 month at 2 - 8°C.
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Biotin-Conjugate
Diluent
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Streptavidin-HRP
Diluent
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Wash Buffer
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Substrate Solution
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Stop Solution
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Once the standard is reconstituted it must be used immediately and cannot be stored for repeated use.
【INTRODUCTION】
Complement C3 is the major plasma proteins of the immune system complement pathways. The synthesis of these proteins is increased in response to inflammation and infection but at a slower rate than for traditional acute phase proteins. Complement protein C3 is the fourth component to attach in the complement reaction sequence. It is a beta-globulin with a sedimentation 4 coefficient of 5.5 and a molecular weight of 185,000. C3 is a central molecule in the complement system whose activation is essential for all the important functions performed by this system. C3 can promote phagocytosis during an inflammatory response against pathogens, but unregulated activation of C3 could lead to host cell damage. Low level of serum C3 associates with hypocomplementaemia, primary biliary cirrhosis.
【PRINCIPLE OF THE ASSAY】
This assay employs a two-site sandwich ELISA to quantitate COMPLEMENT 3 in Salmon serum, plasma. An antibody specific for COMPLEMENT 3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any COMPLEMENT 3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for COMPLEMENT 3 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of COMPLEMENT 3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
【LIMITATIONS OF THE PROCEDURE】
※ FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
※ The kit should not be used beyond the expiration date on the kit label.
※ Do not mix or substitute reagents with those from other lots or sources.
※ It is important that the Calibrator Diluent selected for the standard curve be
consistent with the samples being assayed.
※ If samples generate values higher than the highest standard, dilute the samples with the appropriate Calibrator Diluent and repeat the assay.
※ Any variation in standard diluent, operator, pipetting technique, washing
technique, incubation time or temperature, and kit age can cause variation in binding.
※ This assay is designed to eliminate interference by soluble receptors,
binding proteins, and other factors present in biological samples. Until all factors have been tested in the ELISA Kit, the possibility of interference cannot be excluded.
【DETECTION RANGE】
3.12 μg/mL - 200 μg/mL. The standard curve concentrations used for the ELISA’s were 200 μg/mL, 100 μg/mL, 50 μg/mL, 25 μg/mL, 12.5 μg/mL, 6.25 μg/mL, 3.12 μg/mL, 0 μg/mL.
【SENSITIVITY】
The limit of detection of Salmon GAG defined as the analyte concentration resulting in an absorbance significantly higher than that of the dilution medium (mean plus 2 standard deviations) was determined to be 1.56 μg/mL (mean of 6 independent assays).
【SPECIFICITY】
This assay has high sensitivity and excellent specificity for detection of Salmon COMPLEMENT 3. No significant cross-reactivity or interference between Salmon COMPLEMENT 3 and analogues was observed.
Note:
Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between Salmon COMPLEMENT 3 and all the analogues, therefore, cross reaction may still exist.
【PRECISION】
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8%
Inter-Assay: CV<12%
【STABILITY】
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2-8°C, which means 7 days at 37°C equaling 12 months at 2-8°C).
Note:
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
【SAMPLE COLLECTION AND STORAGE】
- Serum Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2-8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
- Plasma Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2-8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
- Other biological fluids Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at
-20°C or -80°C. Avoid repeated freeze/thaw cycles.
Note:
- Samples to be used within 5 days may be stored at 2-8°C, otherwise samples must be stored at -20°C (≤ 1 month) or -80°C (≤ 2 months) to avoid loss of bioactivity and contamination.
Sample hemolysis will influence the result, so hemolytic specimen cannot
be detected.
- When performing the assay, bring samples to room temperature.
【SAMPLE PREPARATION】
Salmon serum or plasma samples require no dilution before test. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments.
Manual
* Fro research use only