Mesothelin ELISA Kit

Catalog No. ABIN2859340

Human Mesothelin ELISA Kit Colorimetric assay for quantification of Human Mesothelin.

Quick Overview for Mesothelin ELISA Kit (ABIN2859340)

Target: Mesothelin (MSLN)

Binding Specificity: AA 296-580

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156-10.000 pg/mL

Application: ELISA

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

Product Details Mesothelin ELISA Kit

Minimum Detection Limit: 156 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Mesothelin

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: E296-G580

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: NSO
Immunogen sequence: E296-G580

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:

Tissue Specificity: Expressed in lung. Expressed at low levels in heart, placenta and kidney. Expressed in mesothelial cells. Highly expressed in mesotheliomas, ovarian cancers, and some squamous cell carcinomas (at protein level). .

Plate: Pre-coated

Protocol: human Mesothelin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Mesothelin has been precoated onto 96-well plates. Standards(NSO, E296-G580) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Mesothelin 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 human Mesothelin 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 human Mesothelin 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 human cell culture supernates, serum, plasma(heparin, EDTA), saliva or urine to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human Mesothelin standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 0.98, Standard deviation: 0.044, CV(%): 4.5
• Sample 2: n=16, Mean(ng/ml): 2.73, Standard deviation: 0.098, CV(%): 3.6
• Sample 3: n=16, Mean(ng/ml): 5.72, Standard deviation: 0.235, CV(%): 4.1,
• Sample 1: n=24, Mean(ng/ml): 1.09, Standard deviation: 0.073, CV(%): 6.7
• Sample 2: n=24, Mean(ng/ml): 3.16, Standard deviation: 0.167, CV(%): 5.3
• Sample 3: n=24, Mean(ng/ml): 5.95, Standard deviation: 0.381, CV(%): 6.4

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 Mesothelin

Target: Mesothelin (MSLN)

Alternative Name: MSLN

Background:

Protein Function: Membrane-anchored forms may play a role in cellular adhesion.

Background: Mesothelin, also known as MSLN or MPF is a protein that in humans is encoded by the MSLN gene. This gene is mapped to 16p13.3. This gene encodes a precursor protein that is cleaved into two products, megakaryocyte potentiating factor and mesothelin. Megakaryocyte potentiation factor functions as a cytokine that can stimulate colony formation in bone marrow megakaryocytes. Mesothelian is a glycosylphosphatidylinositol-anchored cell-surface protein that may function as a cell adhesion protein. This protein is overexpressed in epithelial mesotheliomas, ovarian cancers and in specific squamous cell carcinomas.

Synonyms: Mesothelin,CAK1 antigen,Pre-pro-megakaryocyte-potentiating factor,Megakaryocyte-potentiating factor,MPF,Mesothelin, cleaved form,MSLN,MPF,

Full Gene Name: Mesothelin

Cellular Localisation: Cell membrane, Lipid-anchor, GPI-anchor. Golgi apparatus.

Gene ID: 10232

UniProt: Q13421

Pathways: EGFR Signaling Pathway, Positive Regulation of Peptide Hormone Secretion, Intracellular Steroid Hormone Receptor Signaling Pathway, Steroid Hormone Mediated Signaling Pathway, Carbohydrate Homeostasis, cAMP Metabolic Process, Regulation of G-Protein Coupled Receptor Protein Signaling, Positive Regulation of Endopeptidase Activity, Regulation of Carbohydrate Metabolic Process