C5A Kit ELISA

N° du produit ABIN1979368

Détail du produit C5A Kit ELISA

Antigène: C5A (C5a) (Complement Component 5a (C5a))

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 1.5-400 pg/mL

Seuil minimal de détection: 1.5 pg/mL

Application: ELISA

Fonction: Mouse C5a ELISA Kit for cell culture supernatants, plasma, and serum samples.

Type d'échantillon: Serum, Plasma, Cell Culture Supernatant

Analytical Method: Quantitative

Specificité: This ELISA kit shows no cross-reactivity with the following cytokines tested: Mouse BLC, CD27, CD30 Ligand, Eotaxin, Eotaxin-2, Fas Ligand, G-CSF, GM-CSF, ICAM-1, IFN-gamma, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12 p40, IL-13, IL-15, IL-17, IL-21, KC, Leptin, LIX, MCP-1, MCP-5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, PF-4, RANTES, TARC, TCA-3, TNF alpha, TNFRI, VEGF.

Réactivité croisée (Details): This ELISA kit shows no cross-reactivity with the following cytokines tested: Mouse BLC, CD27, CD30 Ligand, Eotaxin, Eotaxin-2, Fas Ligand, G-CSF, GM-CSF, ICAM-1, IFN-gamma, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12 p40, IL-13, IL-15, IL-17, IL-21, KC, Leptin, LIX, MCP-1, MCP-5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, PF-4, RANTES, TARC, TCA-3, TNF alpha, TNFRI, VEGF.

Attributs du produit:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Ingrédients:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Matériel non inclus:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Information d'application

Indications d'application: Recommended Dilution for serum and plasma samples100 - 1,000 fold for serum and EDTA plasma, 10 fold for citrate/heparin plasma.

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Préparation des réactifs:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. 2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants. Suggested dilution for normal serum/EDTA plasma: 100-1,000 fold. Suggested dilution for normal citrate/heparin plasma: 10 fold. The Mouse C5a (Complement Component C5a) ELISA Kit Protocol 3 * Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator. 3. Assay Diluent B should be diluted 5-fold with deionized or distilled water before use. 4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture supernates) into Item C vial to prepare a 50 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 8 µL of the C5a standard (50 ng/ml) from the vial of Item C, into a tube with 992 µL Assay Diluent A or 1x Assay Diluent B to prepare a 400 pg/ml standard solution. Pipette 300 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the 400 pg/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/ml). 8 µL standard + 992 µL 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 400 160 64 25.6 10.24 4.10 1.64 0 pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml 5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to yield 400 mL of 1x Wash Buffer. The Mouse C5a (Complement Component C5a) ELISA Kit Protocol 4 6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure. 7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 200-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 60 µL of HRP-Streptavidin concentrate into a tube with 12 mL 1x Assay Diluent B to prepare a 200-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.

Procédure de l'essai:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or overnight at 4 °C with gentle shaking. 3. Discard 4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking. 5. Discard the solution. Repeat the wash as in step 3. 6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking. 7. Discard the solution. Repeat the wash as in step 3. 8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking. 9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Calcul des résultats:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.

Précision du teste: Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: -20 °C

Stockage commentaire: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Date de péremption: 6 months

Références pour C5A Kit ELISA (ABIN1979368)

Kang, Tsolmon, Mann, Zheng, Zhao, Zhao, Volk, Lokesh, Morris, Gupta, Razaq, Rui, Suh, Gorenstein, Tanaka: "Safety evaluation of intravenously administered mono-thioated aptamer against E-selectin in mice." dans: Toxicology and applied pharmacology, Vol. 287, Issue 1, pp. 86-92, (2015) (PubMed).

Détail du antigène

Antigène: C5A (C5a) (Complement Component 5a (C5a))

Autre désignation: C5a (C5a Produits)

Synonymes: C5a Kit ELISA, C5b Kit ELISA, CPAMD4 Kit ELISA, Hc Kit ELISA, RGD1561905 Kit ELISA, complement C5 Kit ELISA, C5 Kit ELISA

ID gène: 15139

UniProt: P06684

Pathways: Système du Complément, Carbohydrate Homeostasis