C4BPA ELISA Kit (Complement Component 4 Binding Protein, alpha) ELISA Kit
- Sandwich ELISA
- 1.56 ng/mL - 100 ng/mL
- Untere Nachweisgrenze
- 1.56 ng/mL
- The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of C4BPa in rat serum, plasma.
- Plasma, Serum
- Analytische Methode
- This assay has high sensitivity and excellent specificity for detection of C4 Binding Protein Alpha (C4BPa)
- 0.65 ng/mL
- 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
- TMB Substrate
- Stop Solution
- Wash Buffer (30 x concentrate)
- Instruction manual
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.
- 100 μL
- 3 h
- Prepare all reagents, samples and standards,
- Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
- Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
- Aspirate and wash 3 times,
- Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
- Aspirate and wash 5 times,
- Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
- Add 50μL Stop Solution. Read at 450nm immediately.
- Aufbereitung der Reagenzien
- Bring all kit components and samples to room temperature (18-25 °C) before use.
- Standard - Reconstitute the Standard with 1.0 mL 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 200 ng/mL. Please firstly dilute the stock solution to 100 ng/mL and the diluted standard serves as the highest standard (100 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.12 ng/mL, 1.56 ng/mL, and the last EP tubes with Standard Diluent is the blank as 0 ng/mL.
- 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).
- 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).
- TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
- Making serial dilution in the wells directly is not permitted.
- Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
- 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.
- The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
- If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
- Contaminated water or container for reagent preparation will influence the detection result.
- Aufbereitung der Proben
- It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
- If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
- If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
- Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%
- Nur für Forschungszwecke einsetzbar
- The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
- 4 °C/-20 °C
- Informationen zur Lagerung
- For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
- For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.
- 6 months
- Andere Bezeichnung
- C4 Binding Protein Alpha (C4BPa) (C4BPA ELISA Kit Abstract)
- CRES, c4bp, C4BP, PRP, complement component 4 binding protein, alpha chain, C4b-binding protein alpha chain, complement component 4 binding protein alpha, complement component 4 binding protein, alpha, LOC419851, c4bp, C4BPA, C4bpa