Complement C4 ELISA Kit

Produktnummer ABIN6974428

Produktdetails Complement C4 ELISA Kit

Target: Complement C4 (C4) (Complement 4 (C4))

Reaktivität: Ratte

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 12.5 ng/mL - 800 ng/mL

Untere Nachweisgrenze: 12.5 ng/mL

Applikation: ELISA

Verwendungszweck: For the quantitative determination of rat complement 4 (C4) concentrations in serum, plasma, tissue homogenates.

Proben: Plasma, Serum, Tissue Homogenate

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of rat C4. No significant cross-reactivity or interference between rat C4 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between rat C4 and all the analogues, therefore, cross reaction may still exist.

Sensitivität: 3.12 ng/mL

Bestandteile:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Probenmenge: 100 μL

Testdauer: 1 - 4.5 h

Plattentyp: Pre-coated

Protokoll:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Aufbereitung der Reagenzien:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer (1x) - If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 800 ng/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undi luted Standard serves as the high standard (800 ng/mL). Sample Diluent serves as the zero standard (0 ng/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. 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).

Note:

Recommend to dilute the serum or plasma samples with Sample Diluent (1:1000) before test. The suggested 1000-fold dilution can be achieved by adding 5 μL sample to 95 μL of Sample Diluent first, then complete the 1000-fold dilution by adding 5 μL of this solution to 245 μL of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments.

Testpräzision: Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Haltbarkeit: 6 months

Referenzen für Complement C4 ELISA Kit (ABIN6974428)

Liu, Chen, Yang, Jiang, Chen, Tu, Mao, Hu, Gan, Li, Li: "iTRAQ-based proteomic analysis to identify the molecular mechanism of Zhibai Dihuang Granule in the Yin-deficiency-heat syndrome rats." in: Chinese medicine, Vol. 13, pp. 2, (2018) (PubMed).

Details zu Complement C4

Target: Complement C4 (C4) (Complement 4 (C4))

Andere Bezeichnung: Complement 4,C4 (C4 Produkte)

Synonyme: C4B1 ELISA Kit, C4B12 ELISA Kit, C4B2 ELISA Kit, C4B3 ELISA Kit, C4B5 ELISA Kit, C4BD ELISA Kit, C4B_2 ELISA Kit, C4F ELISA Kit, CH ELISA Kit, CO4 ELISA Kit, CPAMD3 ELISA Kit, C4 ELISA Kit, C4A2 ELISA Kit, C4A3 ELISA Kit, C4A4 ELISA Kit, C4A6 ELISA Kit, C4AD ELISA Kit, C4S ELISA Kit, CPAMD2 ELISA Kit, RG ELISA Kit, C4-1 ELISA Kit, C4b ELISA Kit, C4-2 ELISA Kit, Slp ELISA Kit, Ss ELISA Kit, complement C4B (Chido blood group) ELISA Kit, complement C4A (Rodgers blood group) ELISA Kit, complement component 4A (Rodgers blood group) ELISA Kit, complement 4 ELISA Kit, complement component 4A ELISA Kit, complement component 4B (Chido blood group) ELISA Kit, C4B ELISA Kit, C4A ELISA Kit, C4a ELISA Kit, C4 ELISA Kit, C4b ELISA Kit

Hintergrund:

Abbreviation: C4

Alias: N/A

UniProt: P08649

Pathways: Komplementsystem