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应用
特点和优势
- 适于少量(1 mg)和大量(5 mg)偶联物。
- 完整水相法 - 不需要DMF
- 从过量的FITC快速凝胶过滤分离偶联物
- 偶联物和F/P比测定完整实验方案
- 试剂足够至少进行5次5mg蛋白质的偶联反应，可供在扩大化前确定最佳F/P比。
- 应用和实验方案参考
原理
分析说明
1.在碳酸盐-重碳酸盐缓冲液中溶解蛋白质和FITC。
2.边搅拌边缓慢将FITC加入蛋白质。盖上箔片，在室温下搅拌2小时。
3.在G-25柱上将自由FITC和偶联物分开。收集组分。
4.收集含有偶联物的组分。
5.分光光度法确定F/P偶联物比例。
6.用1%牛血清白蛋白和0.1%叠氮钠稳定，0-5℃储存。
法律信息
仅试剂盒组分
- Sephadex G-25 column, 3.5 mL 1
- Sephadex G-25 column, 9.1 mL 1
- Fluorescein isothiocyanate isomer I - F7250KC-2MG 2 mg
试剂盒组分也可单独购买
- P3813Phosphate buffered saline, powder, pH 7.4, for preparing 1 L solutions 5 pkg
警示用语：
Warning
危险声明
预防措施声明
危险分类
Eye Irrit. 2
储存分类代码
10 - Combustible liquids
闪点(°F)
Not applicable
闪点(°C)
Not applicable
历史批次信息供参考:
分析证书(COA)
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
Can Product FITC1, FluoroTag™ FITC Conjugation Kit, be used with antibodies at concentrations less than 5 mg/mL?
You can scale down the reaction for labeling. A possible problem occurs in using the column to remove the free FITC. If the volume is too small, the column cannot be used. We would recommend using dialysis to remove the free (unconjugated) FITC.
Can the columns in Product FITC1, FluoroTag™ FITC Conjugation Kit, be reused?
Wash the column with 35 ml (10 X column volumes) of PBS to remove unbound fluorophore. This is sufficient to regenerate the column. For prolonged storage, wash the column with 10 mL of PBS containing 0.05% sodium azide and store capped at 2-8 °C, with about 1 mL buffer above the gel.
When using Product FITC1, FluoroTag™ FITC Conjugation Kit, should I adjust the dye:protein ratio for the size of the protein being labeled?
Adjustments for dye/protein ratios should only be made for very small proteins, i.e. < 25 kDa. Here, you should use less protein per dye portion. In many cases, this weight amount of protein is IgG with MW 150 kDa, and the kit would provide a molar excess of dye to provide attachment of a few dyes per IgG. If one wants to attempt labeling of a very small protein using the same weight ratio, there is a good chance there would not be enough dye/protein.
My antibody is in solution containing azide. Can I use the antibody directly in the reaction when using Product FITC1, FluoroTag™ FITC Conjugation Kit?
It is not recommended to have azide in the reaction mix. If the buffer contains amines or sodium azide, dialyze protein solution (1 mL) against PBS, pH 7.4 (approximately 1000 mL), overnight at 2-8 °C.
How do I find price and availability?
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What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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