Clone | F4/1 |
Isotype | IgG1 |
Product Type | Monoclonal Antibody |
Units | 100 µg |
Host | Mouse |
Application | ELISAFlow CytometryImmunohistochemistryWestern Blotting |
BackgroundThe F4/1 antibody recognizes both the free and protein-conjugated (either soluble or cell bound) form of fluorescein or carboxyfluorescein. Sensitivity of the FITC-anti- FITC system is reportedly similar to that of the biotin-avidin system. When conjugated to biotin or enzyme-labeled, this antibody is also suitable for the conversation of fluorescein-labeled detection into immunoenzymatic assays.
Source
Immunogen: F4/1 is a hybridoma formed by fusing SP-2/0 Agl4 cells with spleen cells of a BALB/c mouse that had been immunised with FITC-KLH.
Product
Product Form: FITC
Formulation: Provided as solution in phosphate buffered saline with 0.08% sodium azide
Purification Method: Protein A/G Chromatography
Concentration: See vial for concentration
ApplicationsThis antibody can be used for Western blotting (1-2 µg/ml), immunohistochemistry (frozen and formalin/paraffin tissues, 2-4 µg/ml) and flow cytometry.Optimal concentration should be evaluated by serial dilutions.
Functional Analysis: Western Blotting
Positive Control: Antibodies labeled with FITC
StorageProduct should be stored at -20ºC. Aliquot to avoid freeze/thaw cycles
Product Stability: Products are stable for one year from purchase when stored properly
Shipping Conditions: Ship at ambient temperature, freeze upon arrival
CautionThis product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Nordic-MUbio accepts no liability for any inaccuracies or omissions in this information.
References1. Balogh P; et al. 'Hapten mediated identification of cell membrane antigens using an anti-FITC monoclonal antibody.' J. Immunol. Methods 1994, 169, 35-40.
Safety Datasheet(s) for this product:NM_Sodium Azide/wp-content/uploads/SDS/Antibody SDS with Sodium Azide Noridic-MUbio.pdf
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英 ['aɪsətaɪp]美 ['aɪsəˌtaɪp]
n. 同型动物(或植物),图形文字,象征性图像;同位型;同模;同号模式
同型对照(阴性对照)管:A同型+B同型+C同型
单阳性对照
A检测管:A+B同型+C同型
B检测管:B+A同型+C同型
C检测管:C+A同型+B同型
不存在荧光重叠的两个抗体不需要调节补偿。
样本为PBMC,想用三色标记Treg细胞,分别为CD4-PECY,CD25-Fitc,Foxp3-PE,想知道如何设置荧光补偿,用的是BD的流式仪,1、是设置阴性管一个,单阳性管每种一个,样品管一个么?
荧光素强度依次为PE, PECY5 APC FITC, PE最强,FITC最弱。 所以,建议抗体组合为:
Foxp3-PE,CD4-FITC,CD25-PECy5.5或者CD25-APC。
同时购买三个相应的同型对照抗体。
样品管设置如下:
1. cell + FITC-isotype + Pecy5.5( or APC)-isotype + PE-isotype
2. Cell + FITC-isotype + CD25-mAb + PE-isotype
3. Cell + PE-mAb + FITC-isotype + Pecy5.5( or APC)-isotype
4. Cell + CD4-FITC + Foxp3-PE + CD25 -PECy5.5( or APC)。 [这一管的数据可以拿来分析用]。
在充分了解调整电压和补偿原理的基础上,也可以做一些简化。如果不太懂,就按照这个来。其中Foxp3的抗体和同型对照抗体都是在固定破膜以后再加,不是与CD4或者CD25同时加。
比如你用anti-MHC-II,mouse IgG1-FITC,那同型就用非特异性的mouse IgG1-FITC,同样浓度
手头上,有eBioscience家的同型抗体,为PE标记(mouseIgG1,k),说明书写使用为2.5ug/test。
按照说明书的浓度加入后结果发现,同型对照抗体组比实验组和阴性对照组(不表达CD80)荧光强度要高,很明确我的同型对照加入的剂量多了,但正确的加法是多少?按照质量算吗?请高手指教,非常感谢!!
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