可用的 EMSA 试剂盒超过 424 个，可用于多数广泛研究的转录因子。
简单—识别 DNA 结合蛋白质
经济—一体化系统！
灵敏度高—基于 HRP 的检测系统
安全—无放射性
Panomics EMSA（凝胶迁移分析）试剂盒用于识别与 DNA 相互作用的蛋白质十分有效。 这种快捷的技术基于自由 DNA 和 DNA/蛋白质复合体在自然（非变性）聚丙烯酰胺凝胶中不同的电泳移动性。 过程很简单：
1. 将生物素标记的 DNA 探针和待研究的蛋白质（提纯或原始提取）中混合温育。
2. 将混合物在非变性聚丙烯酰胺凝胶上分离。
3. 观察与蛋白质/DNA 复合体对应的漂移色带。
与其他商业化 EMSA 试剂盒不同的是，Panomics 的试剂盒包含了特定转录因子的探针。 我们目前供应的 150 个试剂盒与蛋白质/DNA 阵列 I 和 II 上的转录因子对应。 此外，我们还提供包含任意三个探针组的组合件。 探针组也可以单独购买。
 | Panomics NFkB EMSA Kit.Lane 1: Labeled EMSA Probe only with no sample, Lane 2: Labeled EMSA Probe with untreated sample. Lane 3: Labeled EMSA Probe with treated sample. Lane 4: Treated sample with cold and labeled EMSA probes.. | |
要确认某个蛋白质的 DNA 结合活性是否明确，您只需将过量未标记的 DNA 探针加入到探针与细胞核提取物的混合物中。 冷探针会与标记的 DNA 探针竞争，以结合到蛋白质上。 结果，色带的强度将会变弱或消失。
Panomics Electrophoretic-Mobility Shift Assay (EMSA) Kits are useful tools for identifying proteins that interact with DNA. This rapid technique is based on the separation of free DNA from protein/DNA complexes due to the differences in their electrophoretic mobility in native (non-denaturing) polyacrylamide gels. When a protein binds specifically to a labeled dsDNA sequence, it migrates slower than non-bound dsDNA in a polyacrylamide gel, thus resulting in discrete bands corresponding to the individual protein/DNA complex. A typical EMSA experiment is performed by incubating a biotin-labeled transcription factor (TF) Probe with treated and untreated nuclear extracts. The protein/DNA complexes are separated on a non-denaturing polyacrylamide gel. The gel is transferred to a nylon membrane and detected using strepatvidin-HRP and a chemiluminescent substrate. The shifted bands corresponding to the protein/DNA complexes are visualized relative to the unbound dsDNA. The bands are visualized after exposure to film or chemiluminescent-imaging system.
| Panomics NFkB EMSA Kit.Lane 1: Labeled EMSA Probe only with no sample, Lane 2: Labeled EMSA Probe with untreated sample. Lane 3: Labeled EMSA Probe with treated sample. Lane 4: Treated sample with cold and labeled EMSA probes.. |
To confirm whether the DNA binding activity of a particular protein is specific, you simply add an excess of cold, unlabeled DNA probe to the mixture of probe and nuclear extract. The cold probe competes with the labeled DNA probe for binding to the protein. As a result, the band's intensity is reduced or eliminated.
For more specific binding, you may wish to add unlabeled specific dsDNA probe (cold probe; provided) to the protein/DNA reaction mixture which competes with the labeled dsDNA probe (biotin-labeled probe) for binding to the protein. This causes the labeled probe to migrate to the bottom of the gel and reduces the intensity of the shifted band.
Panomics EMSA Kits are suitable for validation of results obtained using Procarta Transcription Factor Plex Assay, Protein/DNA Arrays (Cat. #s MA1010 to MA1015) or to validate binding activity of a specific transcription factor (protein) to DNA. Enough reagents are provided to perform 25 binding reactions. Panomics' TF EMSA Kits measure the activity of specific transcription factors in nuclear extracts. The assay is highly specific, precise, and requires a minimum of 5 ug of protein/well. Panomics currently offers more than 400 EMSA assays.
Panomics EMSA Kits are ideal for validating the results obtained using Protein/DNA Arrays. You can also use them to assess the DNA binding activity of a specific transcription factor.
All-in-one kit
Each EMSA Kit is supplied with:
| —TF* probe | —5X Binding Buffer |
| —Cold (unlabeled) TF* probe | —Loading Dye |
| —Streptavidin-HRP Conjugate | —Poly d(I-C) |
| —HRP Detection Reagents | —Distilled H2O |
| —Control Nuclear Extract | —2X Blocking Buffer |
| —Control Probe | —10X Wash Buffer |
| —Cold (unlabeled) Control Probe | —Enough reagents are provided in each kit to perform 25 gel-shift assays. |
