Anti·His Antibodies、BSA-free

RGS·HisまたはHHHHエピトープのあるHisタグタンパク質の高感度な検出のために

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RGS·His Antibody, BSA-free (100 µg)

Cat. No. / ID: 34650

100 µg マウス抗·RGS(H)₄（凍結乾燥、BSAフリー、1000 mL使用液に対して）

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$943.00

Tag

RGS·His

Tetra·His

Penta·His

Quantity

Anti·His Antibodies、BSA-freeは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

C末端、N末端および内在性Hisタグの特異性が高い検出
RGS·Hisエピトープの高感度で高度に特異的な検出
HHHHHエピトープの高感度で高度に特異的な検出

Product Details

RGS·His Antibody、BSA-free、Tetra·His Antibody、BSA-freeおよびPenta·His Antibody、BSA-freeのようなマウス抗·Hisモノクローナル抗体（5xHis抗体および4xHis抗体）を使用して、Hisタグのある遺伝子組み換えタンパク質を検出します。これらの変異体のそれぞれが異なるHisタグエピトープを検出します。すべてのQIAGENマウスモノクローナル抗体と同様に血清フリーの条件下で調製され、ウイルス、マイコプラズマ、混入免疫グロブリンが存在しないことを保証します。

Performance

QIAexpress Anti·His Antibodies（5xHis抗体および4xHis抗体）を使用し、Hisタグを介して多くの異なるタンパク質が検出されました（表 “AntiHis Antibodiesの性能”を参照）。このパネルのタンパク質の検出における3種類のAnti·His Antibodiesの感度については、図 “QIAexpress Anti·His Antibodiesの感度”を参照してください）。Anti·His Antibodiesは、コロニーブロッティングによる陽性発現コロニーの同定（図 “コロニーブロッティングによる陽性発現コロニーの同定”を参照）、およびHisタグチオレドキシンの検出（図 “Penta·His Antibodyにより酵母細胞内で検出されたHisタグチオレドキシン”を参照）を可能にします。抗体は、免疫組織染色法での使用にも最適です（図 “Penta·His Antibodiesを使用した免疫組織染色”を参照）。

Anti∙His Antibodiesの性能

特徴	RGS·His Antibody、BSA-free	Tetra·His Antibody、BSA-freeおよびPenta·His Antibody、BSA-free
検出	N末端Hisタグ（エピトープRGSHHHHを含む）	N末端、C末端および内在性4xHis（Tetra·His Antibody、BSA-free）または5xHisタグ（Penta·His Antibody、BSA-free)
検出したエピトープ	RGSHHHH	HHHH（Tetra·His Antibody、BSA-free）またはHHHHH（Penta·His Antibody、BSA-free）
ドットブロットでの感度*	0.5 ng	0.5 ng
ウェスタンブロットでの感度*	2 ng	2 ng
アイソタイプ	マウスIgG1	マウスIgG1
交差反応	E. coli、酵母、哺乳類または昆虫の粗細胞溶解物では無視できるほどわずか	E. coli、酵母、哺乳類または昆虫の粗細胞溶解物では無視できるほどわずか
形式	凍結乾燥	凍結乾燥
ワーキングソリューション	1000 mL	1000 mL

* 色素原基質を使用した検出。感度は、タンパク質によって異なる場合があります。

QIAexpress Anti∙His Antibodiesで検出されたタンパク質

	タンパク質	タグの位置	配列
A	DHFR	N末端	MRGSHHHHHHGSVRPLNCIV..........
B	DHFR	C末端	.........VYEKKDGSRSHHHHHH
C	チオレドキシン	N末端	MRGSHHHHHHGSSDKIIHLT..........
D	TNF-α	N末端	MRHHHHHHQSTESMIRDV.........
E	TFIIAγ	N末端	MGSSHHHHHHSSGLVPRGSH.....
F	シャペロニン	C末端	..........AKIAKDGSRSHHHHHH
G	DNAポリメラーゼ	C末端	..........EDWLSAKERSHHHHHH
H	TFIIAαß	内在性	......QQQHQPQQQQHHHHHHHQQAQPQQTVP....

See figures

抗His抗体の感度

コロニーブロッティングによる陽性発現クローンの同定。

酵母細胞でPenta·His Antibodyにより検出したHisタグチオレドキシン。

Penta·His Antibodiesで免疫組織染色。

Principle

QIAexpress Systemは、発現、精製、検出のための材料と、6xHisタグタンパク質のアッセイを提供します。検出用には、QIAexpressマウスモノクローナル抗体が、最高の純度と活性を保証する条件下で産生されます。ハイブリドーマ細胞培養は無血清条件で行うので、調製物はウイルス、マイコプラズマ、混入免疫グロブリンがないことを保証します。吸着クロマトグラフィーによる精製は全体が生理的pHで行われるため、自然抗体が最高の割合となる調製物ができます。

Procedure

QIAexpress、検出およびアッセイハンドブックは、すべてのQIAexpress検出およびアッセイ製品に付属しており、アッセイのの設計および実行のための詳細なプロトコール例およびガイドラインが含まれます。

Applications

RGS·His Antibody、BSA-free、Tetra·His Antibody、BSA-freeおよびPenta·His Antibody、BSA-freeを含む、QIAexpress Detection Systemsは、Hisタグタンパク質の効率的な検出を可能にし、以下を含む多くのアプリケｰションに最適です。

コロニー、ドット、ウェスタンプロッティング法
陽性発現クローンのスクリーニング
Hisタグタンパク質の発現レベルと安定性のモニタリング
免疫沈降およびELISA
免疫細胞染色および免疫組織染色

Supporting data and figures

抗His抗体の感度

抗His抗体による6xHisタグタンパク質の検出（中央がTetra-His抗体）. A: DHFR、B: DHFR、C: チオレドキシン、D: TNF-α、E: TFIIAγ、F:シャペロニン、G DNAポリメラーゼ、H: TFIIAαß、検出されたタグの位置と配列については、表「QIAexpress Anti·His Antibodiesで検出されたタンパク質」を参照。示した量の精製6xHisタグタンパク質を、ニトロセルロース膜に添加し、1/2000に希釈した示した抗His一次抗体で検出を行い、次にAP標識ウサギ抗マウスIgGおよびNBT/BCIPで発色検出した。

Resources

Download Safety Data Sheets for QIAGEN product components.

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

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

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

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

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

Anti·His Antibody epitopes and dissociation constants

Antibody	Epitope	Dissociation constant K_d (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 K_d is dependent on the individual 6xHis-tagged protein.

FAQ ID -385

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

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

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

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

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