产品英文名称：Mouse IFN-γ ELISPOT Pair

产品中文名称：小鼠干扰素-γ Elispot 对

Mouse IFN-γ ELISPOT Pair 产品介绍

The enzyme-linked immunospot (ELISPOT) assay is a powerful tool for detecting and enumerating individual cells that secrete a particular protein in vitro. Based on the sandwich ELISA, the ELISPOT assay derives its specificity and sensitivity by employing high affinity capture and detection antibodies and enzyme-amplification. Although originally developed for analyzing specific antibody-secreting cells, the assay has been adapted for measuring the frequencies of cells that produce and secrete other effector molecules, such as cytokines. The sensitivity of the assay lends itself to measurement of even very low frequencies of cytokine producing cells (e.g., 1/300,000). Unique strengths of the assay include high sensitivity, high throughput, high content analysis, minimal volume of biological material required, applicability to frozen/thawed biological samples, and compatibility with other assays. For example, cells analyzed by ELISPOT can be transferred for cloning, proliferation assays, flow cytometry, or other methods of analysis.

This product contains sufficient reagents for five 96-well plates, including unlabelled anti-cytokine capture antibody (no azide/low endotoxin format), biotinylated anti-cytokine detection antibody and a Certificate of Analysis that provides lot-specific optimal reagent concentrations.

小鼠干扰素-γ Elispot 对的保存方法

Store undiluted at 4° C and protected from prolonged exposure to light. Do not freeze.

Mouse IFN-γ ELISPOT 对的推荐使用步骤

A. BD™ ELISPOT Streptavidin Horseradish Peroxidase (Cat. No. 557630) is recommended.

B. Coating Buffer (1X Phosphate Buffered Saline : 8 g NaCl; 0.2 g KCl; 1.44 g Na2HPO4-7H2O, 0.24 gKH2PO4; dissolved in H2O to a final volume of 1 liter. Adjust pH to 7.2, sterile-filter (0.2 μm-sized pore filter) and store at 4°C.

C. Blocking Solution: complete tissue culture medium (e.g. RPMI 1640 containing 10% Fetal Bovine Serum [FBS] and 1% Penicillin-Streptomycin.L-Glutamine [Gibco-BRL No. 10378-016]).

D. Wash Buffer I: 1X PBS containing 0.05% Tween-20 (0.5 mL Tween-20 per 1 L PBS).

E. Wash Buffer II: 1X PBS

F. Dilution Buffer: 1X PBS containing 10% FBS.

G. BD™ ELISPOT AEC Substrate Set (Cat. No. 551951) is recommended. Alternatively, Prepare as follows:

1. Prepare AEC (3-Amino-9-ethyl-carbazole; Sigma A-5754) stock solution: mix 100 mg AEC in 10 mL DMF (N,NDimethylformamide; Sigma D-4551). Caution: dispense DMF in fume hood. Store solution in glassware.

2. Prepare 0.1 M Acetate Solution: add 148 mL of 0.2 M acetic acid/glacial acidic acid to 352 mL of 0.2 M sodium acetate. Adjust volume to 1L DI water; adjust pH to 5.0.

3. For Final Substrate Solution, add 333.3 μL of AEC stock solution to 10 mL 0.1 M Acetate Solution. Filter through 0.45 μm filter. Add 5 μLof H 202 (30%) and use immediately.

Mouse IFN-γ ELISPOT Pair 的优越性

•高灵敏度

•高通量，高含量的分析

•最小体积的生物材料的要求

•适用于冷冻/解冻的生物样品 •与其他检验方法兼容。例如，细胞分析。BD™ ELISPOT可以转移克隆，增殖实验，流式细胞术或其他分析方法。

Mouse IFN-γ ELISPOT Pair 使用注意事项

1. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.

2．Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosi