Search
Disclaimer
Clicking the images or links will redirect you to a website hosted by BenchSci that provides third-party scientific content. Neither the content nor the BenchSci technology and processes for selection have been evaluated by us; we are providing them as-is and without warranty of any kind, including for use or application of the Thermo Fisher Scientific products presented.
Invitrogen
Penta-His Tag Monoclonal Antibody
{{$productOrderCtrl.translations['antibody.pdp.commerceCard.promotion.promotions']}}
{{$productOrderCtrl.translations['antibody.pdp.commerceCard.promotion.viewpromo']}}
{{$productOrderCtrl.translations['antibody.pdp.commerceCard.promotion.promocode']}}: {{promo.promoCode}} {{promo.promoTitle}} {{promo.promoDescription}}. {{$productOrderCtrl.translations['antibody.pdp.commerceCard.promotion.learnmore']}}
图: 1 / 7
Penta-His Tag Antibody (P-21315) in WB
Detection of an oligohistidine fusion protein with the Penta-His mouse monoclonal antibody. Twofold serial dilutions of an Escherichia coli lysate containing overexpressed oligomycin sensitivity-conferring protein (OSCP) fused with a hexahistidine domain were run an SDS-polyacrylamide gel and blotted onto a PVDF membrane. The blot was stained with the SYPRO Ruby protein blot stain (Product # S-11791) to detect the entire protein profile (top). After imaging, the blot was incubated with Penta-His mouse IgG1 monoclonal antibody (P... View More
产品信息
P-21315
应用
建议稀释比
已发表文章
产品规格
种属反应
Tag
已发表种属
Tag
宿主/亚型
Mouse
/ IgG1
分类
Monoclonal
类型
Antibody
抗原
Histidine Tag (5xHis)
偶联物
形式
Lyophilized
纯化类型
purified
保存液
PBS with PEG, sucrose
内含物
0.08% sodium azide
保存条件
4°C
运输条件
Ambient (domestic); Wet ice (international)
RRID
产品详细信息
Sufficient material is supplied to stain 50-100 minigel blots (8 cm × 10 cm). For immunodetection of oligohistidine fusion proteins on blotting membranes, use the Penta-His antibody at a final concentration of between 0.1 µg/mL and 0.2 µg/mL (1:1,000 to 1:2,000 dilution of the stock solution). Standard buffers and blocking agents, such as 3% BSA (bovine serum albumin) may be used; however, milk powder should not be used, as it reduces sensitivity. Denaturation of the proteins is recommended to ensure exposure of the oligohistidine domain. Detect the Penta-His antibody with an antimouse IgG secondary antibody conjugated to either alkaline phosphatase or horseradish peroxidase. The enzyme activity can then be visualized using a chromogenic, fluorogenic or chemiluminescent substrate.
Upon receipt, the lyophilized antibody should be stored at 4°C. When properly stored, this product is stable for at least one year. To prepare a stock solution, reconstitute the antibody in 0.5 mL of deionized water (dH2O) to a final concentration of 0.2 mg/mL. Store the solution for up to 3 months at 4°C. For storage up to 6 months, divide the solution into single-use aliquots and freeze at -20°C. AVOID REPEATED FREEZING AND THAWING.
Immunohistochemical methods can be used to investigate the location of recombinant proteins expressed in particular cells within tissues or organs, as well as for determination of the subcellular localization of proteins after targeting to particular cell compartments. The Penta-His antibody has been used to detect a hexahistidine aggrecan fusion protein in the perinuclear Golgi and the endoplasmic reticulum of transiently transfected CHO cells. The technique can also be used to localize cellular proteins that interact with hexahistidine fusion proteins. Preparation of the cells or tissue: Fix the tissue with 2% paraformaldehyde and permeabilize with 0.25% Triton™ X-100. This relatively mild fixation protocol best preserves epitopes and best maintains the position of the protein within the cellular structure. Immunodetection for cells and tissues: Before applying the antibody to the tissue, block nonspecific binding sites using 5% bovine serum albumin (BSA). For immunodetection of the oligohistidine fusion protein, use the Penta-His antibody at a final concentration of 410 µg/mL (1:20 to 1:50 dilution of the stock solution). Detect the Penta-His antibody with an anti-mouse IgG secondary antibody conjugated to a fluorescent dye, alkaline phosphatase, or horseradish peroxidase. The fluorescent dye can be seen directly using a fluorescence-based microscope. The enzyme activity can be detected using a chromogenic or fluorogenic substrate.
靶标信息
The oligohistidine domain is a Ni2+-binding peptide sequence comprising a string of four to six histidine residues. When the DNA sequence corresponding to the oligohistidine domain is fused in frame with a gene of interest, the resulting fusion protein can be easily purified using a nickel-chelating resin. The Penta-His monoclonal IgG1 antibody provides a sensitive method for specific detection of fusion proteins that have an oligohistidine domain comprising five or six consecutive histidine residues. The antibody does not recognize tetrahistidine domains or domains in which the histidine string is interrupted by another amino acid. The antibody binds to the oligohistidine domain regardless of the surrounding amino acid context and even when the group is partially hidden, although subtle differences in the amino acid context may change the sensitivity limit for a particular fusion protein. This antibody can be used to detect oligohistidine fusion proteins on western blots, dot blots or colony blots, or in tissues.
仅用于科研。不用于诊断过程。未经明确授权不得转售。
