CoraLite® Plus 750-conjugated 6*His, His-Tag Monoclonal antibody
6*His, His-Tag Monoclonal Antibody for WB
Host / Isotype
Mouse / IgG1
Reactivity
recombinant protein
Applications
WB
Conjugate
CoraLite® Plus 750 Fluorescent Dye
CloneNo.
1B7G5
验证数据展示
经过测试的应用
| Positive WB detected in | Transfected HEK-293T cells |
推荐稀释比
| Application | Dilution |
|---|---|
| Western Blot (WB) | WB : 1:1000-1:4000 |
| 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
CL750-66005 targets 6*His, His-Tag in WB applications and shows reactivity with recombinant protein samples.
| Tested Applications | WB Application Description |
| Tested Reactivity | recombinant protein |
| Immunogen | Peptide 种属同源性预测 |
| Host / Isotype | Mouse / IgG1 |
| Class | Monoclonal |
| Type | Antibody |
| Full Name | 6*His, His-Tag |
| Synonyms | |
| Calculated Molecular Weight | 1 kDa |
| Gene Symbol | |
| Gene ID (NCBI) | |
| Conjugate | CoraLite® Plus 750 Fluorescent Dye |
| Excitation/Emission Maxima Wavelengths | 755 nm / 780 nm |
| Form | Liquid |
| Purification Method | Protein G purification |
| Storage Buffer | PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3. |
| Storage Conditions | Store at -20°C. Avoid exposure to light. 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 | |
|---|---|
| WB protocol for CL Plus 750 6*His, His-Tag antibody CL750-66005 | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |