ADAM12 ELISA Kit

Catalog No. ABIN1889434

Product Details ADAM12 ELISA Kit

Target: ADAM12 (ADAM Metallopeptidase Domain 12 (ADAM12))

Binding Specificity: AA 29-513

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156-10.000 pg/mL

Minimum Detection Limit: 156 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse ADAM12

Brand: PicoKine™

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin)

Analytical Method: Quantitative

Specificity: Expression system for standard: CHO
Immunogen sequence: R29-S513

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

Sensitivity: <10pg/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: CHO
Immunogen sequence: R29-S513

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.

Comment:

Sequence similarities: Contains 1 disintegrin domain.

Tissue Specificity: Expressed during early developing mesenchymal cells that give rise to skeletal muscle, bones and visceral organs. Not expressed in adult normal muscle but expressed in regenerating muscle. .

Plate: Pre-coated

Protocol: mouse ADAM12 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for ADAM12 has been precoated onto 96-well plates. Standards(CHO, R29-S513) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for ADAM12 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 ADAM12 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 10,000pg/mL, 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312pg/mL, 156pg/mL mouse ADAM12 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 supernatants, serum or plasma(heparin) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse ADAM12 standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 1.45, Standard deviation: 0.062, CV(%): 4.3
• Sample 2: n=16, Mean(ng/ml): 3.76, Standard deviation: 0.139, CV(%): 3.7
• Sample 3: n=16, Mean(ng/ml): 6.87, Standard deviation: 0.24, CV(%): 3.5,
• Sample 1: n=24, Mean(ng/ml): 1.54, Standard deviation: 0.089, CV(%): 5.8
• Sample 2: n=24, Mean(ng/ml): 3.81, Standard deviation: 0.171, CV(%): 4.5
• Sample 3: n=24, Mean(ng/ml): 7.02, Standard deviation: 0.365, CV(%): 5.2

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 ADAM12

Target: ADAM12 (ADAM Metallopeptidase Domain 12 (ADAM12))

Alternative Name: ADAM12 (ADAM12 Products)

Background:

Protein Function: Involved in skeletal muscle regeneration, specifically at the onset of cell fusion. Also involved in macrophage-derived giant cells (MGC) and osteoclast formation from mononuclear precursors.

Background: ADAM12(Disintegrin and metalloproteinase domain-containing protein 12), also known as MLTN, is an enzyme that in humans is encoded by the ADAM12 gene. This gene encodes a member of the ADAM (a disintegrin and metalloprotease) protein family. Members of this family are membrane-anchored proteins structurally related to snake venom disintegrins, and have been implicated in a variety of biological processes involving cell-cell and cell-matrix interactions, including fertilization, muscle development, and neurogenesis. This gene has two alternatively spliced transcripts: a shorter secreted form and a longer membrane-bound form. The shorter form is found to stimulate myogenesis. By RT-PCR and immunoblot analyses that expression of mouse Adam12 increases during muscle regeneration, while the levels of other ADAMs remain constant. Immunofluorescence analysis revealed staining of small, newly formed muscle fibers in regenerating but not normal adult muscle cells. By using of fluorescence in situ hybridization, the ADAM12 gene is mapped to human chromosome 10q26.3.

Synonyms: Disintegrin and metalloproteinase domain-containing protein 12,ADAM 12,3.4.24.-,Meltrin-alpha,Adam12,Mltna,

Full Gene Name: Disintegrin and metalloproteinase domain-containing protein 12

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

Gene ID: 11489

UniProt: Q61824

Pathways: EGFR Signaling Pathway