Clone | 15D4 |
Isotype | IgG1 |
Product Type | Monoclonal Antibody |
Units | 100 µg |
Host | Mouse |
Species reactivity | BovineHumanRat |
Application | Enzyme ImmunoassayImmunoprecipitationWestern Blotting |
BackgroundSNAPs (soluble NSF attachment proteins), acting in concert with SNAREs (SNAP receptors) and the N-ethylmaleimide-sensitive fusion protein (NSF), are required for the fusion of transport vesicles to their target membranes in synaptictransmission, intra-Golgi transport, endosome-to-endosome fusion and transcytotic vesicles-to-plasma membrane transport. Vesicle-to-target membrane docking (initial contact) occurs when the vesicle SNARE binds to its cognate target membrane SNARE. Beta-SNAP (or SNAP in brain) then binds to this docking complex and mediates the binding of NSF and thus the formation of a 20 S fusion particle. It is thought that, once NSF is bound, ATP hydrolysis by NSF initiates the fusion process. Whereas alpha-SNAP is expressed in all mammalian tissues, beta-SNAP is expressed only in brain.
Synonyms: alpha/beta SNAP, mouse anti alpha soluble NSF attachment protein,SNAPA
SourceMonoclonal antibody 15D4 obtained immunization of Balb/Cmice withrecombinanthuman alpha -SNAP
Immunogen: Balb/Cmicewereimmunizedwithrecombinanthuman alpha -SNAP.The resultant hybridomas were selected for reactivity with theimmunogenby ELISAand with the antigenbyWesternblot.
Product100 µg monoclonal antibody in 0.02M sodium phosphate, pH 7.5, 0.15M sodium chloride, 50% glycerol. No preservatives have been added.
Product Form: Unconjugated
Formulation: 0.02M sodium phosphate, pH 7.5, 0.15M sodium chloride, 50% glycerol. No preservatives have been included in the antibody solution.
Purification Method: Theantibodywaspurifiedto>95%homogeneitybystandard chromatographic techniques
Concentration: See vial for concentration
SpecificityHuman alpha -SNAP
ApplicationsOptimal concentration should be evaluated by serial dilutions. Clone 4E4 anti-?SNAP specifically recognizes ?-SNAP as a single band of ~36 kDa on western blot of rat kidney, rat brain and MDBK cells and does not recognize ?-SNAPApplication: A dilution of 0.5-5 ug/ml is suggested for western blot and 2-10 ug/ml for immunoprecipitation.
Functional Analysis: Western Blotting
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: Room Temperature
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.
Protein Reference(s)
Database Name: UniProt
Accession number: P54920 (Human)
Species Accession: Human
Safety Datasheet(s) for this product:NM_Glycerol/wp-content/uploads/SDS/SDS Antibodies with glycerol Nordic-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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