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Invitrogen
HisG Epitope Tag Monoclonal Antibody, HRP
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HisG Epitope Tag Antibody (R941-25) in WB
Western 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 detecte... View More
产品信息
R941-25
产品规格
种属反应
Tag
已发表种属
Tag
宿主/亚型
Mouse
/ IgG2a
分类
Monoclonal
类型
Antibody
抗原
6x His synthetic peptide: -His-His-His-His-His-His-Gly
偶联物
形式
Liquid
纯化类型
purified
保存液
PBS
内含物
no preservative
保存条件
Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID
产品详细信息
NOTE: 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.
靶标信息
Epitope 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. HisG antibodies allow detection of recombinant proteins containing a polyhistidine sequence: His-His-His-His-His-His-Gly (6xHis-Gly epitope). HisG antibodies can be used to detect expression of fusion proteins from bacterial, insect, and mammalian cells.
仅用于科研。不用于诊断过程。未经明确授权不得转售。
