Ccl12 ELISA Kit

Catalog No. ABIN8092870

Mouse Ccl12 ELISA Kit, Colorimetric assay for quantification of Mouse Ccl12.

Quick Overview for Ccl12 ELISA Kit (ABIN8092870)

Target: Ccl12 (Chemokine (C-C Motif) Ligand 12 (Ccl12))

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 7.81 pg/mL - 500 pg/mL

Application: ELISA

Sample Type: Plasma, Serum

Grade: MiniSample

Product Details Ccl12 ELISA Kit

Minimum Detection Limit: 7.81 pg/mL

Purpose: Small Sample Mouse CCL12/MCP-5 (Monocyte Chemotactic Protein 5) ELISA Kit

Analytical Method: Quantitative

Cross-Reactivity (Details): This kit recognizes Mouse CCL12/MCP-5 in samples.No significant cross-reactivity or interference between Mouse CCL12/MCP-5 and analogues was observed

Sensitivity: 3.97 pg/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (50 x concentrate)
• HRP Conjugate (50 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Application Details

Sample Volume: 25 μL

Assay Time: 3.5 h

Plate: Pre-coated

Protocol:

1. Add 25 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 50 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 50 µL HRP Conjugate Working Solution. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 50 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 25 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.

Reagent Preparation:

1. Equilibrate all reagents to room temperature (18-25 °C) before use. If the kit will not be used up in one assay, please only take out the necessary strips and reagents for present experiment, and store the remaining strips and reagents at required condition.
2. Centrifuge the standard at 10,000x g for 1 min. Reconstitute with 1 mL of the Standard & Sample Diluent, allow to stand for 10 minutes, and gently invert several times until completely dissolved. Alternatively, vortex briefly at low speed, remove bubbles by low-speed centrifugation if necessary.This produces a 500 pg/mL stock solution. Prepare serial dilutions according to experimental requirements. Recommended dilution gradient: 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 0 pg/mL Serial Dilution Procedure: Dispense 500 μL of Standard & Sample Diluent into 7 EP tubes, respectively. Transfer 500 μL of the stock solution into the first tube. Sequentially transfer 500 μL from the preceding tube into the next tube for subsequent dilutions. Designate the last tube as the blank, do not transfer any solution into it. Aliquot the stock and store at -20 ° C. Use within two weeks and avoid repeated freeze-thaw cycles. Serially diluted standards should be prepared immediately, prior to use.
3. Biotinylated Detection Ab working solution: Calculate the required amount before the experiment (50 μL/well) plus a slight excess Centrifuge the Concentrated Biotinylated Detection (50x) Ab at 800x g for 1 min. Dilute to 1x with Biotinylated Detection Ab Diluent at a ratio of 1:49. Prepare the working solution immediately, prior to use.
4. Calculate the required amount before the experiment (50 μL/well) plus a slight excess. Centrifuge the Concentrated HRP Conjugate (50x) at 800 x g for 1 minute. Dilute to 1x with HRP Conjugate Diluent at a ratio of 1:49. Prepare the working solution immediately, prior to use.
5. Wash Buffer: Prepare the working Wash Buffer by diluting 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water yielding 750 mL. Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved. For same day use only.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of 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 the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.

Assay Precision:

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level of the target were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level of the target were tested on 3 different plates, 20 replicates in each plate.

Both intra-CV and inter-CV are < 10%.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials, so they are stable up to 12 months after receipt of the kit. The Reference Standard, Biotinylated Detection Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2. For used kit: When the kit is used, the remaining reagents 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 zip-seal the foil pouch.

Expiry Date: 12 months

Target Details for Ccl12

Target: Ccl12 (Chemokine (C-C Motif) Ligand 12 (Ccl12))

Alternative Name: Monocyte Chemotactic Protein 5

Background:

Target Synonym: CCL12,Mcp5,Scya12,C-C motif chemokine 12,MCP-1-related chemokine,Monocyte chemoattractant protein 5,Monocyte chemotactic protein 5 (MCP-5),Small-inducible cytokine A12

UniProt: Q62401

Pathways: Cellular Response to Molecule of Bacterial Origin