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详细说明

6*His, His-Tag Monoclonal antibody

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产品编号 66005-1-Ig
  • 商品说明

经过测试的应用

Positive WB detected inrecombinant protein
Positive IP detected inTransfected HEK-293 cells
Positive IF detected inTransfected HEK-293 cells
Planning an IF experiment? We recommend our CoraLite® Plus 488, CoraLite®594 and CoraLite® Plus 647 conjugated versions of this antibody.

推荐稀释比

APPLICATIONDILUTION
Western Blot (WB)WB : 1:5000-1:50000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for IP and 1:5000-1:50000 for WB
Immunofluorescence (IF)IF : 1:200-1:800
Sample-dependent, check data in validation data gallery

产品信息

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

Tested ApplicationsIF, IP, WB, ELISA
Cited ApplicationsCell Treatment, ChIP, CoIP, ELISA, FC, IF, IHC, IP, WB
Tested ReactivityRecombinant Protein
Cited ReactivityHuman, Mouse
ImmunogenPeptide 种属同源性预测
Host / IsotypeMouse / IgG1
ClassMonoclonal
TypeAntibody
Full Name6*His, His-Tag
Synonyms6*His, 6*His, His Tag, 6*His, His-Tag, 6histag, 6XHis, HHHHHH, HIS, His probe, His Tag, His-probe, HISTIDINE TAGGED
Calculated molecular weight1 kDa
Gene symbol
Gene ID (NCBI)
RRIDAB_11232599
ConjugateUnconjugated
FormLiquid
Purification MethodProtein A purification
Storage BufferPBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore 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.



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