Anti·His Antibodies, BSA-free

用于具有 RGS·His 或 HHHH 表位的 His 标签蛋白的灵敏性检测

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RGS·His Antibody, BSA-free (100 µg)

Cat. No. / ID:  34650

100 µg 小鼠抗·RGS(H)4(冻干,不含 BSA,用于 1000 mL 工作溶液)
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Tag
RGS·His
Tetra·His
Penta·His
Anti·His Antibodies, BSA-free 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • C-末端、N-末端和内部 His 标签的高度特异性检测
  • RGS·His 表位的高度灵敏性和特异性检测
  • HHHHH 表位的高度灵敏性和特异性检测

Product Details

单克隆小鼠抗 His 抗体(5xHis 抗体和 4xHis 抗体),例如 RGS·His Antibody, BSA-free、Tetra·His Antibody, BSA-free 和 Penta·His Antibody, BSA-free,用于检测携带 His 标签的重组蛋白。这三种变体各检测一种不同的 His 标签表位。像所有 QIAGEN 小鼠单克隆抗体一样,它们都是在无血清条件下制备的,从而确保没有病毒、支原体和污染性免疫球蛋白。

Performance

使用 QIAexpress 抗 His 抗体(5xHis 抗体和 4xHis 抗体),可通过其 His 标签检测许多不同的蛋白质(参见表格“抗 His 抗体的性能”)。关于这三种抗 His 抗体在检测这组蛋白质方面的灵敏度,请参见图片  “QIAexpress 抗 His 抗体的灵敏度”)。抗 His 抗体能够通过菌落印迹法鉴定阳性表达克隆(参见图片 “通过菌落印迹法鉴定阳性表达克隆”)以及检测 His 标签硫氧还蛋白(参见图片 “通过 Penta·His 抗体检测酵母细胞中的 His 标签硫氧还蛋白”)。这些抗体也非常适合用于免疫组织化学分析(参见图片 “使用 Penta·His 抗体进行免疫组织化学分析”)。

抗 His 标签抗体的性能

特点 RGS·His Antibody, BSA-free Tetra·His Antibody, BSA-free 和 Penta·His Antibody, BSA-free
检测 N-末端 His 标签(包含 RGSHHHH 表位) N-末端、C-末端和内部 4xHis (Tetra·His Antibody, BSA-free) 或 5xHis (Penta·His Antibody, BSA-free) 标签
检测到的表位 RGSHHHH HHHH (Tetra·His Antibody, BSA-free) 或 HHHHH (Penta·His Antibody, BSA-free)
斑点印迹分析中的灵敏度* 0.5 ng 0.5 ng
蛋白质印迹分析中的灵敏度* 2 ng 2 ng
同种型 小鼠 IgG1 小鼠 IgG1
交叉反应 与天然 E. coli、酵母、哺乳动物或昆虫细胞裂解物的交叉反应可忽略不计 与天然 E. coli、酵母、哺乳动物或昆虫细胞裂解物的交叉反应可忽略不计
剂型 冻干 冻干
工作溶液 1000 mL 1000 mL

使用 QIAexpress 抗 His 抗体检测的蛋白质

  蛋白质 标签位置 序列
A DHFR N-末端 MRGSHHHHHHGSVRPLNCIV..........
B DHFR C-末端 .........VYEKKDGSRSHHHHHH
C 硫氧还蛋白 N-末端 MRGSHHHHHHGSSDKIIHLT..........
D TNF-α N-末端 MRHHHHHHQSTESMIRDV.........
E TFIIAγ N-末端 MGSSHHHHHHSSGLVPRGSH.....
F 伴侣蛋白 C-末端 ..........AKIAKDGSRSHHHHHH
G DNA 聚合酶 C-末端 ..........EDWLSAKERSHHHHHH
H TFIIAαß 内部 ......QQQHQPQQQQHHHHHHHQQAQPQQTVP....

 

See figures

Principle

QIAexpress System 提供了用于 6xHis 标签蛋白表达、纯化和检测的材料。用于检测的 QIAexpress 小鼠单克隆抗体是在保证具有最高纯度和活性的条件下生产的。在无血清条件下进行杂交瘤细胞培养,以确保制剂不含病毒、支原体和污染性免疫球蛋白。完全在生理 pH 下通过吸附层析进行纯化,从而产生具有最高天然抗体比例的制剂。

Procedure

随所有 QIAexpress 检测产品一起提供的《QIAexpress 检测手册》包含用于设计和执行检测的详细示例方案和指南。

Applications

QIAexpress Detection System,包括 RGS·His Antibody, BSA-free、Tetra·His Antibody, BSA-free 和 Penta·His Antibody, BSA-free,可高效检测 His 标签蛋白,适合许多应用,包括:

  • 菌落、斑点和蛋白质印迹分析
  • 阳性表达克隆的筛选
  • 监测 His 标签蛋白的表达水平和稳定性
  • 免疫沉淀和 ELISA
  • 免疫细胞化学和免疫组织化学

Supporting data and figures

Resources

安全数据表 (3)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (1)
For Anti·His Antibodies, Anti·His HRP Conjugates, Penta·His™ Alexa Fluor® Conjugates, Penta·His Biotin Conjugate, Ni-NTA Conjugates, Tag·100™ Antibody, Streptavidin–R-PE, 6xHis Protein Ladder, Ni-NTA HisSorb™ Strips and Plates 
Certificates of Analysis (1)
Kit Handbooks (1)
For Anti·His Antibodies, Anti·His HRP Conjugates, Penta·His™ Alexa Fluor® Conjugates, Penta·His Biotin Conjugate, Ni-NTA Conjugates, Tag·100™ Antibody, Streptavidin–R-PE, 6xHis Protein Ladder, Ni-NTA HisSorb™ Strips and Plates 
Supplementary Protocols (1)

Publications

Localization of the gD-binding region of the human herpes simplex virus receptor, HveA.
Whitbeck JC; Connolly SA; Willis SH; Hou W; Krummenacher C; Ponce de Leon M; Lou H; Baribaud I; Eisenberg RJ; Cohen GH;
J Virol; 2001; 75 (1):171-80 2001 Jan PMID:11119586
Multiple sulfatase deficiency is caused by mutations in the gene encoding the human C(alpha)-formylglycine generating enzyme.
Dierks T; Schmidt B; Borissenko LV; Peng J; Preusser A; Mariappan M; von Figura K;
Cell; 2003; 113 (4):435-44 2003 May 16 PMID:12757705
Screening for soluble expression of recombinant proteins in a 96-well format.
Knaust RK; Nordlund P;
Anal Biochem; 2001; 297 (1):79-85 2001 Oct 1 PMID:11567530
A human cDNA expression library in yeast enriched for open reading frames.
Holz C; Lueking A; Bovekamp L; Gutjahr C; Bolotina N; Lehrach H; Cahill DJ;
Genome Res; 2001; 11 (10):1730-5 2001 Oct PMID:11591650
Phosphorylation of the vesicle-tethering protein p115 by a casein kinase II-like enzyme is required for Golgi reassembly from isolated mitotic fragments.
Dirac-Svejstrup AB; Shorter J; Waters MG; Warren G;
J Cell Biol; 2000; 150 (3):475-88 2000 Aug 7 PMID:10931861

FAQ

How should I dissolve the lyophilized Anti-His and Tag Antibodies?
Dissolve the lyophilized Anti-His Antibodies (100 µg) in 500 µl water per vial. The final concentration is 0.2 mg/ml. Dissolve the Anti-His Antibody Selector Kit antibodies (3 µg) in 15 µl water per tube (final concentration, 0.2 mg/ml).
-100
What epitopes do the Anti-His Conjugates and Anti-His Antibodies recognize?

The table below lists the epitopes recognized by the Anti-His Conjugates and Anti-His Antibodies. 'H' designates a histidine residue, while 'X' designates any amino acid other than histidine. Note that individual 6xHis-tagged proteins are often recognized better by one Anti-His Antibody than by the others, possibly because of subtle differences in the exact conformation of the 6xHis tag and other parts of the protein in the vicinity of the tag. To choose the optimal antibody for your specific 6xHis-tagged protein and application, QIAGEN offers the Anti-His Antibody Selector Kit. The kit includes 3 µg of each of the three antibodies for direct comparison and economical selection of the one that gives the best results.

Epitopes recognized by QIAexpress Anti-His HRP Conjugates and Anti-His Antibodies


Peptide Penta-His Tetra-His RGS-His
XHHHHHHX
+ +
XXHHHHHX
+ +
XHXHHHHX
+
XHHHHXHX
+
XRGSHHHHX
+ +
XRGSHHHHHHX
+ + +
XHXHHXHX

Note that the presence of specific residues N- or C-terminal to the epitope is not required.

FAQ ID -170
Which Anti-His Antibody is the most sensitive for my protein of interest?

Penta-His and Tetra-His Antibody bind to proteins that have 6xHis tags regardless of the surrounding amino-acids, even when the tag is partially hidden. Subtle differences in the antigen recognition sites of these antibodies mean that some 6xHis-tagged proteins are detected with higher sensitivity by one Anti-His Antibody than by the others (see figure below). Therefore, to achieve optimum results, we recommend comparison of these antibodies in parallel for new applications and proteins. QIAGEN offers the Anti-His Antibody Selector Kit for this purpose.


 

Sensitivity of QIAexpress Anti·His Antibodies



 

Note:  RGS His Antibody regonizes the RGS(His)4 epitope that is found on proteins endcoded by most pQE vectors as well as several pRSETan pBlue BacHis -vectors from Invitrogen.

FAQ ID -172
What are the dissociation constants for the Anti-His Antibodies?

Anti·His Antibody epitopes and dissociation constants

Antibody Epitope Dissociation constant Kd (M)*
RGS·His Antibody RGSHHHH 1 x 10–8 – 5 x 10–8
Penta·His Antibody HHHHH 5 x 10–8 – 1 x 10–9
Tetra·His Antibody HHHH 1 x 10–8 – 5 x 10–8

* Dissociation constants were measured using surface plasmon resonance (BIACORE) technology. The exact value of Kd is dependent on the individual 6xHis-tagged protein.

FAQ ID -385
Are QIAGEN monoclonal Anti His Antibodies glycosylated?
Yes, QIAGEN antibodies are glycosylated to a low extent. We did not determine whether this is N-glycosylation or O-glycosylation. Furthermore, we do not know, if the glycosylation pattern does change, since we do not measure it. Glycosylation does not have any impact onto the function of the antibody. If customers want to use the glycosylchains for coupling reactions we know that this is very difficult to do.
FAQ ID -334
How is an antibody epitope determined?
This is done via epitope mapping. The antibody in question is first tested against different protein fragments and peptides. Once the amino acids detected by the antibody are determined, the surrounding amino acids can be exchanged to reveal the influence of the environment on the detection of a specific epitope.
FAQ ID -593
What blocking reagents do you recommend for chemiluminescent detection using Penta-, Tetra-, and RGS-His antibodies?

Please do not use buffer containing milk powder to block the Western Blot. This will reduce sensitivity. QIAGEN recommends use 3% BSA in TBS-buffer to block the membrane and for incubation of the Anti His Antibody on the membrane. For the binding of the secondary Antibody BSA does not sufficiently block and therfore 10% milk powder in TBS-buffer should be used at this step only. Alternatively, if alkali-soluble casein (Merck, Cat. No. 1.02241) is available in your country it can be used as a blocking reagent throughout the chemiluminescent detection protocol. Please see Table 9 of the QIAxpress Detection and Assay Handbook for additional information.

FAQ ID -1093
What is the difference between BSA-free and BSA-containing RGS-His Antibody?

BSA stabilizes the RGS-His Antibody. We recommend to use the BSA-containing RGS-His Antibody for all standard procedures like dot blots, western blots, etc.. In cases where BSA may interfere with the application, e.g. coupling reactions, the RGS-His Antibody, BSA-free is recommended.

The Tetra-His and Penta-His Antibodies are stable without BSA, and therefore are only offered BSA-free.

FAQ ID -569
How should I store Anti·His and Tag·100 Antibodies?
QIAGEN Anti·His Antibodies should be stored lyophilized until they are to be used. They can be stored lyophilized for 1 year at 2–8°C. In solution they can be stored for 3 months at 2–8°C or for up to 6 months in aliquots at –20°C. Avoid repeated freezing and thawing.
FAQ ID -194