6*His, His-Tag Monoclonal antibody

6*His, His-Tag Monoclonal Antibody for WB, IF/ICC, FC, IP, ELISA
Cat No. 66005-1-Ig

产品说明书

CloneNo. 1B7G5

宿主/亚型

Mouse / IgG1

种属反应性

recombinant protein and More (3)

应用

WB, IF/ICC, FC, IP, ELISA and More (4)

6*His, His Tag, His Tag, 6*His, HHHHHH, 6XHis

缓冲液配方:  PBS and Azide
PBS and Azide
PBS Only
规格: 

-/ -


经过测试的应用

Positive WB detected inrecombinant protein
Positive IP detected inTransfected HEK-293 cells
Positive IF/ICC detected inTransfected HEK-293 cells
Positive FC detected in转染Eg0175 Expi293 cells

推荐稀释比

应用推荐稀释比
Western Blot (WB)WB : 1:5000-1:50000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunofluorescence (IF)/ICCIF/ICC : 1:200-1:800
Flow Cytometry (FC)FC : 0.20 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

产品信息

66005-1-Ig targets 6*His, His-Tag in WB, IHC, IF/ICC, FC, IP, CoIP, ChIP, ELISA applications and shows reactivity with recombinant protein samples.

经测试应用 WB, IF/ICC, FC, IP, ELISA Application Description
文献引用应用WB, IHC, IF, FC, IP, CoIP, ChIP, ELISA
经测试反应性 recombinant protein
文献引用反应性human, mouse, chicken
免疫原 Peptide 种属同源性预测
宿主/亚型 Mouse / IgG1
抗体类别 Monoclonal
产品类型 Antibody
全称 6*His, His-Tag
别名 6*His, His Tag, His Tag, 6*His, HHHHHH, 6XHis
计算分子量 0.84 kDa
基因名称
Gene ID (NCBI)
RRIDAB_11232599
偶联类型 Unconjugated
形式 Liquid
纯化方式Protein G purification
储存缓冲液 PBS with 0.02% sodium azide and 50% glycerol , pH 7.3
储存条件Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

背景介绍

Protein tags are a protein or peptide sequences located either on the C- or N- terminal of the target protein. His-tag is often used for affinity purification and binding assays. Expressed. The His-tag antibody is a useful tool for monitoring of the His-tagged proteins and recognizes His-tags placed at N-terminal, C-terminal, and internal regions of fusion proteins expressed in bacteria, insect, and mammalian cells. A His-tag (polyhistidine tag) consists of at least six histidine residues that are located at the N- or C-terminus of recombinant proteins. It is commonly used for affinity purification and protein binding experiments.

 

The recombinant protein has only 6 histidine residues as a tag. Is this sufficient to be detected by your antibody? Is the His-tag antibody able to detect N-terminal and C-terminal His tags?

The His-tag antibody was raised using a 6-His tag peptide as the immunogen, which is ~1 kDa. Some scientists use more than 6 histidine residues as a tag for recombinant proteins to increase affinity to metal ions. A 6-histidine tag is sufficient for specific recognition by the His-tag antibody. It is able to recognize both N-terminal and C-terminal His tags of recombinant proteins produced in various expression systems, including bacteria, yeasts, insect, and mammalian cells.

 

After purification, I can detect my protein by western blotting using the His-tag antibody but can also see additional bands running lower. What might they represent?

They may represent cleavage products of the recombinant protein. Unspecific protein cleavage can occur during protein production (e.g., in bacteria) or protein purification post the cell lysis steps. Changing expression conditions (temperature, time, induction methods, medium) and using protease inhibitors during purification helps to minimize protein degradation. Additional steps, such as ion exchange, size exclusion, or chromatography, allow isolation of the full-length product.

 

Can a His-tag antibody be used in applications other than western blotting?

Yes, this antibody has been successfully used for immunofluorescence (IF), immunoprecipitation (IP), and indirect ELISA analysis.


实验方案

Product Specific Protocols
IF protocol for 6*His, His-Tag antibody 66005-1-IgDownload protocol
IP protocol for 6*His, His-Tag antibody 66005-1-IgDownload protocol
FC protocol for 6*His, His-Tag antibody 66005-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

发表文章

SpeciesApplicationTitle
WB

Cell

The WDR11 complex is a receptor for acidic-cluster-containing cargo proteins

Authors - Huaqing Deng

Nature

UDP-glucose accelerates SNAI1 mRNA decay and impairs lung cancer metastasis.

Authors - Xiongjun Wang

Nature

IGF1R is an entry receptor for respiratory syncytial virus.

Authors - Cameron D Griffiths
WB

Nature

CRISPR screens unveil signal hubs for nutrient licensing of T cell immunity.

Authors - Lingyun Long
WB

Science

Structural insight into the SAM-mediated assembly of the mitochondrial TOM core complex.

Authors - Qiang Wang
WB

Nature

Fatal swine acute diarrhoea syndrome caused by an HKU2-related coronavirus of bat origin.

Authors - Peng Zhou
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