CD25 ELISA Kit

Catalog No. ABIN1672833

This Mouse CD25 ELISA Kit is a Colorimetric ELISA Kit designed to quantify Mouse CD25.

Quick Overview for CD25 ELISA Kit (ABIN1672833)

Target: CD25 (IL2RA) (Interleukin 2 Receptor, alpha (IL2RA))

Binding Specificity: AA 22-236

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 78-5000 pg/mL

Application: ELISA

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

Product Details CD25 ELISA Kit

Minimum Detection Limit: 78 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse CD25/IL-2sR alpha

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO,E22-K236

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

Sensitivity: <5pg/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: E22-K236

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 CD25 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for CD25 has been precoated onto 96-well plates. Standards (NSO,E22-K236) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD25 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 CD25 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 313pg/mL, 156pg/mL, 78pg/mL mouse CD25 standard solutions into the pre-coated 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, serum or plasma( heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. We recommend that each mouse CD25 standard solution and each sample is measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 564, Standard deviation: 32.7, CV(%): 5.8
• Sample 2: n=16, Mean(pg/ml): 1244, Standard deviation: 75.9, CV(%): 6.1
• Sample 3: n=16, Mean(pg/ml): 2860, Standard deviation: 157.3, CV(%): 5.5,
• Sample 1: n=24, Mean(pg/ml): 758, Standard deviation: 54.6, CV(%): 7.2
• Sample 2: n=24, Mean(pg/ml): 1654, Standard deviation: 134, CV(%): 8.1
• Sample 3: n=24, Mean(pg/ml): 3242, Standard deviation: 217, CV(%): 6.7

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 CD25

Target: CD25 (IL2RA) (Interleukin 2 Receptor, alpha (IL2RA))

Alternative Name: IL2RA

Background:

Protein Function: Receptor for interleukin-2.

Background: CD25 is the alpha chain of the IL-2 receptor. It is a type I transmembrane protein present on activated T cells, activated B cells, some thymocytes, myeloid precursors, and oligodendrocytes that associates with CD122 to form a heterodimer that can act as a high-affinity receptor for IL-2. CD25 has been used as a marker to identify CD4+FoxP3+ regulatory T cells in mice. By in situ hybridization, the IL2R gene was mapped to 10p15-p14. Its soluble form, called sIL-2R may be elevated in these diseases and is occasionally used to track disease progression.

Synonyms: Interleukin-2 receptor subunit alpha,IL-2 receptor subunit alpha,IL-2-RA,IL-2R subunit alpha,IL2-RA,p55,CD25,Il2ra,Il2r,

Full Gene Name: Interleukin-2 receptor subunit alpha

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

Gene ID: 16184

UniProt: P01590

Pathways: JAK-STAT Signaling, Growth Factor Binding, Activated T Cell Proliferation