6X His tag Recombinant Mouse Monoclonal Antibody [A5D1-R] - BSA and Azide free
Catalog# HA610016
6X His tag Recombinant Mouse Monoclonal Antibody [A5D1-R] - BSA and Azide free
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WB
-
IP
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Species independent
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HA601079
含抗保成分
-
unconjugated
概述
产品名称
6X His tag Recombinant Mouse Monoclonal Antibody [A5D1-R] - BSA and Azide free
抗体类型
Recombinant Mouse Monoclonal Antibody
免疫原
Synthetic peptide HHHHHHGCGHHHHHH.
种属反应性
Species independent
验证应用
WB, IP
阳性对照
C-terminal His-tag fusion protein lysates, Mid His-tag fusion protein lysates.
偶联
unconjugated
克隆号
A5D1-R
产品特性
形态
Liquid
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
PBS (pH7.4).
亚型
IgG1
纯化方式
Protein A affinity purified.
应用稀释度
-
WB
-
1:5,000-1:50,000 (C-His)
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WB
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1:1,000-1:5,000 (Mid-His)
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IP
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Use at an assay dependent concentration (C-His)
靶点
功能
His-Tag Antibody is a high quality monoclonal His-Tag antibody (also designated 6X His-Tag antibody or His-Probe antibody) suitable for the detection of the His-Tag protein. His-Tag Antibody is available as both the non-conjugated anti-His-Tag antibody form, as well as multiple conjugated forms of anti-His-Tag antibody, including agarose, HRP, PE, FITC and multiple Alexa Fluor® conjugates. Plasmid vectors for the expression of coding regions of eukaryotic genes in bacterial, insect and mammalian hosts are in common usage; such expression vectors are frequently used to encode hybrid fusion proteins consisting of a eukaryotic target protein and a specialized region designed to aid in the purification and visualization of the target protein. A system that has proven to be very successful relies on the insertion of a six histidine (His6) sequence in the N-terminus of the encoded protein, allowing for efficient coupling to Ni2+-chelating resins and purification by single step affinity chromatography. This polyhistidine sequence can then be removed by specific cleavage at sites recognized by enzymes such as thrombin or enterokinase, permitting the separation of the target protein from the polyhistidine tag. Visualization of such fusion proteins can be achieved by utilizing antibodies generated against specific peptide sequences downstream from the multiple cloning site.
背景文献
1. Hengen, Paul N (1995). Purification of His-Tag fusion proteins from Escherichia coli. Trends in Biochemical Sciences. 20 (7): 285–6.
2. Hochuli, E.; Döbeli, H.; Schacher, A. (January 1987). New metal chelate adsorbent selective for proteins and peptides containing neighbouring histidine residues. Journal of Chromatography A. 411: 177–184.
别名
6 His epitope tag antibody
Hexa His tag antibody
HHHHHH epitope tag antibody
HHHHHH tag antibody
His tag antibody
Polyhistidine Tag antibody
图片
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Western blot analysis of 6X His tag on different lysates with Mouse anti-6X His tag antibody (HA610016) at different dilutions.
Lane 1/2: C-terminal His-tag fusion protein lysates
Lane 3/4: Mid His-tag fusion protein lysates
Lysates/proteins at 50 ng/Lane.
Observed band size: 30/40 kDa
Exposure time: 30 seconds;
12% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA610016) at different dilutions was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature. -
His-tag was immunoprecipitated in 2µg C-terminal His-tag fusion protein lysate with HA610016. Western blot was performed from the immunoprecipitate using HA610016 at 1/2,000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 60 mins at room temperature.
Lane 1: C-terminal His-tag fusion protein lysate (input).
Lane 2: HA610016 IP in C-terminal His-tag fusion protein lysate.
Lane 3: Mouse IgG instead of HA610016 IP in C-terminal His-tag fusion protein lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"