HisG Epitope Tag Monoclonal Antibody, HRP/HisG标签抗体

货号：R941-25

规格：50 µl

价格：4050

产品类型：标签抗体

品牌：Thermo Fisher

物种：Tag

宿主：小鼠

抗体亚型：其它

荧光染料：HRP

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抗体类型:	单抗	同型对照：	IgG1
免疫原性：	6x His synthetic peptide: -His-His-His-His-His-His-Gly	用法：	1:1000(WB)

Product Specific InformationNOTE: R941-25 was previously sold under product #46-1009. When re-ordering, please use #R941-25.This antibody is designed to specifically detect recombinant proteins containing the seven amino acid 6xHis-Gly epitope. This epitope is present in many expression vectors encoding an N-terminal 6xHis tag. The HisG antibody recognizes the following sequence: -His-His-His-His-His-His-Gly and can be used to detect expression of fusion proteins from bacterial, insect, and mammalian cells.Anti-HisG-HRP Antibody was prepared by crosslinking the Anti-HisG Antibody with horseradish peroxidase using glutaraldehyde. It has been tested in immunoblotting and ELISA procedures. It was tested against purified Positope™ control protein (5 ng). The Positope™ control protein is a 53 kDa recombinant protein that contains seven epitope tags, including His(C-term), HisG, c-Myc, and V5. Low background was observed using chemiluminescent or alkaline phosphatase reagents for detection. In western blot experiments with purified protein, 50 ng (for Anti-HisG-HRP Antibody) of the Positope protein gave a detectable signal.For western blot, dilute in PBS containing 0.05% Tween-20 and 5% nonfat, dry milk (PBSTM). Using chemiluminescence as the detection method, no cross-reactivity has been observed in bacterial lysates. In mammalian lysates, a few cross-reactive proteins have been observed upon overexposure of blots. If you use alkaline phosphatase-conjugated secondary antibody, do not use PBS. Phosphate inhibits alkaline phosphatase. Use Tris-Buffered Saline (TBS) instead. Be sure to wash the western blot or microtiter wells before adding the horseradish peroxidase-conjugated secondary antibody. Azide will inhibit HRP activity. This product contains enough material for 25 western blots; assumes 10 mL buffer per western blot.Background/Target InformationEpitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein-specific antibodies are not available. Expression of recombinant proteins in E. coli as a fusion protein with neighboring histidine residues is one of the most popular methods of epitope tags. The affinity of the histidine-tag motif to Ni2+ by chelation is strong and selective enough to enable purification of the protein to homogeneity by affinity chromatography on a Ni2 +-NTA adsorbent.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

HisG Epitope Tag Antibody (R941-25) in WBWestern blot analysis of HisG was performed by loading 20 µg of whole cell extracts of untransfected HEK-293 cell lysate (lane1) and HEK-293 transiently over expressing HisG-RPS6 (lane2) using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), Xcell SureLock Electrophoresis system (Product # EI0002), Novex sharp Pre-stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for at least 1 hour at room temperature. HisG was detected at ~35 kDa using HisG - HRP Mouse Monoclonal Antibody (Product # R941-25) at a 1:1000 dilution in 2.5 % skim milk incubated for 1 hour on a rocking platform. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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参考文献：

1.PloS oneExploring the Functional Complementation between Grp94 and Hsp90."R94125 was used in western blot to discover structural elements that distinguish Grp94 from Hsp90"AuthorsMaharaj KA,Que NL,Hong F,Huck JD,Gill SK,Wu S,Li Z,Gewirth DT2.PLoS geneticsA systems genetics approach implicates USF1, FADS3, and other causal candidate genes for familial combined hyperlipidemia."R941-25 was used in western blot to identify causal candidate genes for Mexican Familial Combined Hyperlipidemia-related traits"AuthorsPlaisier CL,Horvath S,Huertas-Vazquez A,Cruz-Bautista I,Herrera MF,Tusie-Luna T,Aguilar-Salinas C,Pajukanta P

技术参数

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