CCL3 ELISA Kit

Catalog No. ABIN411322

Mouse CCL3 ELISA Kit Colorimetric assay for quantification of Mouse CCL3.

Quick Overview for CCL3 ELISA Kit (ABIN411322)

Target: CCL3 (Chemokine (C-C Motif) Ligand 3 (CCL3))

Binding Specificity: AA 33-262

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.6-1000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Tissue Homogenate, Serum, Plasma (EDTA)

Product Details CCL3 ELISA Kit

Minimum Detection Limit: 15.6 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse M-CSF

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: K33-E262

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <1pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: E.coli
Immunogen sequence: K33-E262

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Plate: Pre-coated

Protocol: mouse M-CSF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for M-CSF has been precoated onto 96-well plates. Standards(E.coli, K33-E262) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for M-CSF is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse M-CSF amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL mouse M-CSF standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, tissue lysates, serum or plasma (EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse M-CSF standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 92, Standard deviation: 5.06, CV(%): 5.5
• Sample 2: n=16, Mean(pg/ml): 316, Standard deviation: 19.28, CV(%): 6.1
• Sample 3: n=16, Mean(pg/ml): 628, Standard deviation: 40.2, CV(%): 6.4,
• Sample 1: n=24, Mean(pg/ml): 127, Standard deviation: 7.493, CV(%): 5.9
• Sample 2: n=24, Mean(pg/ml): 439, Standard deviation: 27.66, CV(%): 6.3
• Sample 3: n=24, Mean(pg/ml): 685, Standard deviation: 51.4, CV(%): 7.5

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

Target Details for CCL3

Target: CCL3 (Chemokine (C-C Motif) Ligand 3 (CCL3))

Alternative Name: CCL3

Background:

Protein Function: Cytokine that plays an essential role in the regulation of survival, proliferation and differentiation of hematopoietic precursor cells, especially mononuclear phagocytes, such as macrophages and monocytes. Promotes the release of proinflammatory chemokines, and thereby plays an important role in innate immunity and in inflammatory processes. Plays an important role in the regulation of osteoclast proliferation and differentiation, the regulation of bone resorption, and is required for normal bone development. Required for normal male and female fertility. Promotes reorganization of the actin cytoskeleton, regulates formation of membrane ruffles, cell adhesion and cell migration. Plays a role in lipoprotein clearance.

Background: M-CSF, also called CSF1, consists of a 14-amino acid peptide, longer than the usual 8-to-11 mer recognized by most CTLs. The M-CSF gene is mapped to 1p21-p13 and contains 10 exons and 9 introns spanning 20 kb1. Although it is a single-copy gene, its expression results in the synthesis of several mRNAs, ranging in size from about 1.5 to 4.5 kb2. There are 2 forms of M-CSF, with 224 and 522 amino acids, resulting from alternative splicing3. Furthermore, M-CSF has a role in development of the placenta. Uterine M-CSF concentration is regulated by a synergistic action of estradiol and progesterone. M-CSF is produced by uterine glandular epithelial cells. It had been found that FMS, the M-CSF receptor, is expressed in placenta and choriocarcinoma cell lines4.

Synonyms: Macrophage colony-stimulating factor 1,CSF-1,MCSF,Processed macrophage colony-stimulating factor 1,Csf1,Csfm,

Full Gene Name: Macrophage colony-stimulating factor 1

Cellular Localisation: Cell membrane, Single-pass type I membrane protein.

Gene ID: 12977

UniProt: P07141

Pathways: Cellular Response to Molecule of Bacterial Origin, Autophagy