CD40 ELISA Kit

Catalog No. ABIN921090

This Mouse CD40 ELISA Kit is a Colorimetric ELISA Kit designed to quantify Mouse CD40.

Quick Overview for CD40 ELISA Kit (ABIN921090)

Target: CD40

Binding Specificity: AA 24-193

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.6-1000 pg/mL

Application: ELISA

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

Product Details CD40 ELISA Kit

Minimum Detection Limit: 15.6 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse CD40/TNFRSF5

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: V24-R193

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: NSO
Immunogen sequence: V24-R193

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 CD40 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for CD40 has been precoated onto 96-well plates. Standards(NSO, V24-R193) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD40 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 CD40 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 CD40 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, cell lysates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse CD40 standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 95, Standard deviation: 6.46, CV(%): 6.8
• Sample 2: n=16, Mean(pg/ml): 336, Standard deviation: 17.47, CV(%): 5.2
• Sample 3: n=16, Mean(pg/ml): 535, Standard deviation: 26.22, CV(%): 4.9,
• Sample 1: n=24, Mean(pg/ml): 120, Standard deviation: 9.96, CV(%): 8.3
• Sample 2: n=24, Mean(pg/ml): 361, Standard deviation: 23.10, CV(%): 6.4
• Sample 3: n=24, Mean(pg/ml): 541, Standard deviation: 30.30, CV(%): 5.6

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 CD40

Target: CD40

Alternative Name: CD40

Background:

Protein Function: Receptor for TNFSF5/CD40LG. Transduces TRAF6- and MAP3K8-mediated signals that activate ERK in macrophages and B cells, leading to induction of immunoglobulin secretion. .

Background: CD40 is a cell surface receptor that is expressed on the surface of all mature B cells, most mature B-cell malignancies, and some early B-cell acute lymphocytic leukemias, but is not expressed on plasma cells. The CD40 gene is localized to the long arm of chromosome 20, bands q12-q13.2. The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor has been found to be essential in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation.

Synonyms: Tumor necrosis factor receptor superfamily member 5,B-cell surface antigen CD40,Bp50,CD40L receptor,CD40,Cd40,Tnfrsf5,

Full Gene Name: Tumor necrosis factor receptor superfamily member 5

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

Gene ID: 21939

UniProt: P27512

Pathways: NF-kappaB Signaling, Cellular Response to Molecule of Bacterial Origin, M Phase, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Cancer Immune Checkpoints