C3 ELISA Kit

Catalog No. ABIN414464

Product Details C3 ELISA Kit

Target: C3 (Complement Component 3 (C3))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 7.8 ng/mL - 500 ng/mL

Minimum Detection Limit: 7.8 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of C3 in Serum,Plasma,Biological Fluids

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Specificity:

This assay has high sensitivity and excellent specificity for detection of Complement Component 3 (C3).
No significant cross-reactivity or interference between Complement Component 3 (C3) and analogues was observed.

Cross-Reactivity (Details): No significant cross-reactivity or interference between Complement Component 3 (C3) and analogues was observed.

Sensitivity: 2.8 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Application Notes:

• Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
• The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
• Kits from different batches may be a little different in detection range, sensitivity and color developing time.
• Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
• Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
• There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
• Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
• Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
• Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
• Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Complement Component 3 (C3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Complement Component 3 (C3). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Complement Component 3 (C3), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Complement Component 3 (C3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use.
2. Standard - Reconstitute the Standard with 1.0mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 1,000ng/mL. First dilute the stock solution to 500ng/mL and the diluted standard serves as the highest standard (500ng/mL). Prepare 7 tubes containing 0.5mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 500ng/mL, 250ng/mL, 125ng/mL, 62.5ng/mL, 31.2ng/mL, 15.6ng/mL, 7.8ng/mL, and the last EP tubes with Standard Diluent is the blank as 0ng/mL.
3. Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with Assay Diluent A and B, respectively (1:100).
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for once pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Assay Precision:

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 3 (C3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Complement Component 3 (C3) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Handling Advice: The stability of 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.
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.

Storage: 4 °C

Storage Comment:

• For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
• For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
  Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
• For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.

Expiry Date: 6 months

References for C3 ELISA Kit (ABIN414464)

Shen, Zhang, Feng, Chen, Li, Iqbal, Liao, Zhao, Zhai: "iTRAQ-Based Proteomic Analysis Reveals Protein Profile in Plasma from Children with Autism." in: Proteomics. Clinical applications, Vol. 12, Issue 3, pp. e1700085, (2019) (PubMed).

Klupczynska, Swiatly, Hajduk, Matysiak, Dyszkiewicz, Pawlak, Kokot: "Identification of Serum Peptidome Signatures of Non-Small Cell Lung Cancer." in: International journal of molecular sciences, Vol. 17, Issue 4, pp. 410, (2016) (PubMed).

Ma, Cui, Liao, Zhao: "Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease." in: Journal of clinical immunology, Vol. 33, Issue 1, pp. 172-8, (2013) (PubMed).

Burillo, Mateo-Gallego, Cenarro, Fiddyment, Bea, Jorge, Vázquez, Civeira: "Beneficial effects of omega-3 fatty acids in the proteome of high-density lipoprotein proteome." in: Lipids in health and disease, Vol. 11, Issue 1, pp. 116, (2012) (PubMed).

Oberbach, Blüher, Wirth, Till, Kovacs, Kullnick, Schlichting, Tomm, Rolle-Kampczyk, Murugaiyan, Binder, Dietrich, von Bergen: "Combined proteomic and metabolomic profiling of serum reveals association of the complement system with obesity and identifies novel markers of body fat mass changes." in: Journal of proteome research, Vol. 10, Issue 10, pp. 4769-88, (2011) (PubMed).

Kim, Kobashi: "Immobilized arylsulfotransferase." in: Journal of biochemistry, Vol. 102, Issue 3, pp. 487-91, (1988) (PubMed).

Target Details for C3

Target: C3 (Complement Component 3 (C3))

Alternative Name: Complement Component 3 (C3 Products)

Synonyms: AHUS5 ELISA Kit, ARMD9 ELISA Kit, ASP ELISA Kit, C3a ELISA Kit, C3b ELISA Kit, CPAMD1 ELISA Kit, AI255234 ELISA Kit, HSE-MSF ELISA Kit, Plp ELISA Kit, C3 ELISA Kit, C3-1 ELISA Kit, sb:cb26 ELISA Kit, si:dkey-76b14.4 ELISA Kit, wu:fi34b03 ELISA Kit, wu:fj86e11 ELISA Kit, C3d ELISA Kit, asp ELISA Kit, cpamd1 ELISA Kit, c3 ELISA Kit, complement C3 ELISA Kit, complement component 3 ELISA Kit, complement component c3a, duplicate 1 ELISA Kit, complement component 3 L homeolog ELISA Kit, complement component C3 ELISA Kit, complement C3-like ELISA Kit, C3 ELISA Kit, c3a.1 ELISA Kit, c3.L ELISA Kit, LOC443475 ELISA Kit, c3 ELISA Kit, c3-1 ELISA Kit, LOC100060539 ELISA Kit

UniProt: P01024

Pathways: Complement System, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Regulation of G-Protein Coupled Receptor Protein Signaling