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- 文献和实验
- 技术资料
- 免疫原:
None
- 亚型:
IgG1
- 保存条件:
Store at -20°C. Avoid exposure to light. Stable for one year after shipment.
- 克隆性:
Monoclonal
- 标记物:
CoraLite® Plus 488 Fluorescent Dye
- 适应物种:
recombinant protein
- 保质期:
详见产品包装
- 库存:
充足
- 宿主:
Mouse
- 应用范围:
IF/ICC
- 靶点:
6*His, His-Tag
- 抗体英文名:
CoraLite® Plus 488-conjugated 6*His, His-Tag Monoclonal antibody
- 抗体名:
CoraLite® Plus 488-conjugated 6*His, His-Tag Monoclonal antibody
- 规格:
100ul
经过测试的应用
| Positive IF/ICC detected in | Transfected HEK-293 cells |
推荐稀释比
| 应用 | 推荐稀释比 |
|---|---|
| Immunofluorescence (IF)/ICC | IF/ICC : 1:50-1:500 |
| 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
CL488-66005 targets 6*His, His-Tag in IF/ICC applications and shows reactivity with recombinant protein samples.
| 经测试应用 | IF/ICC |
| 文献引用应用 | IF |
| 经测试反应性 | recombinant protein |
| 免疫原 |
fusion protein |
| 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 probe, HIS, HHHHHH, 6XHis |
| 计算分子量 | 0.84 kDa |
| 基因名称 | |
| Gene ID (NCBI) | |
| RRID | AB_2883223 |
| 偶联类型 | CoraLite® Plus 488 Fluorescent Dye |
| 最大激发/发射波长 | 493 nm / 522 nm |
| 形式 | Liquid |
| 纯化方式 | Protein G purification |
| UNIPROT ID | HISTAG |
| 储存缓冲液 | PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3. |
| 储存条件 | 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.
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