- ￥1900 - 6200
- Sino Biological
- 北京
- TB101274
- 2026年01月14日
- 免疫沉淀
相关产品推荐更多 >
万千商家帮你免费找货
0 人在求购买到急需产品
- 详细信息
- 文献和实验
- 技术资料
- 形态：
液体
- 保存条件：
2-8℃
- 克隆性：
其他
- 保质期：
12个月
- 目录编号：
TB101274
- 级别：
免疫学试剂
- 库存：
99
- 供应商：
北京义翘神州科技股份有限公司
- 应用范围：
免疫沉淀
- 抗体英文名：
Immunoprecipitation Kit -DYKDDDDK（Flag®）Tag Immunomagnetic Beads
- 抗体名：
Immunoprecipitation Kit -DYKDDDDK（Flag®）Tag Immunomagnetic Beads
- 规格：
20 Tests/100 Tests
| 规格： | 20 Tests | 产品价格： | ￥1900.0 |
|---|---|---|---|
| 规格： | 100 Tests | 产品价格： | ￥6200.0 |
| Contents | TB101274-20 | TB101274-100 | Storage |
| DYKDDDDK Tag Immunomagnetic Beads1 | 1 mL | 5 mL | 2-8℃ for 12 months |
| NP40 Cell Lysis Buffer2 | 4 mL | 22 mL | -20℃ for 12 months |
| 5×TBST（pH7.4） | Required but not supplied | ||
| 1×TBST（pH7.4） | Required but not supplied | ||
| ddH2O | Required but not supplied | ||
| DYKDDDDK Tag Positive Cell Lysis | 300 μg | 300 μg | -20℃ for 12 months |
| Alkaline Elution Buffer | 3 mL | 15 mL | 2-8℃ for 12 months |
| Acidity Elution Buffer | 3 mL | 15 mL | 2-8℃ for 12 months |
| Neutralization Buffer | 2 mL | 8 mL | 2-8℃ for 12 months |
| DYKDDDDK Synthetic Peptide | Not included (refer related product PP101274) | Not included (refer related product PP101274) | -20℃ for 12 months |
[1] DYKDDDDK Tag Immunomagnetic Beads contain immunomagnetic beads (2 mg/mL) in phosphate buffered saline (PBS, pH 7.4) with sodium azide (0.1%).
[2] Using NP-40 cell lysate buffer in the kit is required,otherwise,the magnetic beads may be precipitated.
The DYKDDDDK Tag Immunomagnetic Beads, conjugated with DYKDDDDK Antibody (101274-MM13), are used for immuneprecipitation (IP) of DYKDDDDK-tagged proteins expressed in vitro expression systems. For IP, the Immunomagnetic Beads are added to a sample containing DYKDDDDK-tagged proteins to form an Immunomagnetic Beads-protein complex. The complex is removed from the solution manually against a Magnetic Separator. The bound DYKDDDDK-tagged proteins are dissociated from the Immunomagnetic Beads using an Elution Buffer.
Antibody: DYKDDDDK (FLAG® epitope Tag) Antibody, Mouse Mab (101274-MM13)
Immunogen: A synthetic peptide corresponding to the DYKDDDDK-tag sequence (CDYKDDDDK).
Clone ID: MM13
Isotype: lgG1
Specificity: Recognize N-terminal and C-terminal Flag Tag in fusion proteins.
Preparation: This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, the synthetic peptide corresponding to the DYKDDDDK-tag sequence (CDYKDDDDK). The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography.
| Items | Lane | |
|
Sample (30 μg) (Whole cell lysate) |
A | B |
| pSTEP2 Transfected 293 | Flag-mFABP4-GST | |
| Beads | SBI Anti-DYKDDDDK (FLAG® epitope Tag) Immunomagnetic Beads-30 μL | |
| WB detection antibody | Anti-DYKDDDDK (FLAG® epitope Tag) Antibody, Mouse MAb (101274-MM13) at 10 μg/mL | |
| Gel | 13% SDS-PAGE reducing gel | |
| Secondary antibody | Dylight 800-labeled antibody to mouse IgG (H+L), at 1:5000 dilution | |
The protocol (Fig. 1) uses 50 μL flag Tag Immunomagnetic Beads, but this can be scaled up or down as required.
Cell Lysis
Cells may be lysed using any standard cell lysis protocol in accordance with your starting materials. We suggest using NP40 Cell Lysis Buffer (supplied with kit). Add prtease inhibitor (such as PMSF at 1mM) if needed.
Immunoprecipitate Target Antigen
1. Add 50 μL of Immunomagnetic Beads into a 1.5 mL microcentrifuge tube.
2. Add 150 μL of 1× TBST buffer to the Immunomagnetic Beads and gently vortex to mix.
3. Place the tube into a Magnetic Separator to collect the Immunomagnetic Beads against the side wall of the tube. Remove and discard the supernatant.
4. Add 1 mL of 1×TBST buffer to the tube. Invert the tube several times or gently vortex to mix for 1 min. Collect the Immunomagnetic Beads with a Magnetic Separator. Remove and discard the supernatant.
5. Add the sample containing target protein (~100 μg of protein in 100 μL) to the pre-washed Immunomagnetic Beads, add 400 μL of 1×TBST buffer and incubate at 37℃ for 20-30 min (or at room temperature for 2h) with mixing.
6. Collect the Immunomagnetic Beads with a Magnetic Separator , remove the unbounded sample and save for analysis.
7. Add 300 μL of 5× TBST buffer to the tube and gently mix. Collect the Immunomagnetic Beads and discard the supernatant. Repeat this wash twice.
8. Add 300 μL of ddH2O to the tube and gently mix. Collect the Beads on a Magnetic Separator and discard the supernatant.
Elute Target Antigen
A. Neutral Elution Protocol
1. Prepare DYKDDDDK peptide (PP101274) at 1mg/mL in PBS.
2. Add 50 μL 1 mg/mL DYKDDDDK peptide to the Immunomagnetic Beads, gently vortex to mix and incubate the sample at 37 ℃ on a rotator for 5-10 min. Elution may be performed at reduced temperatures, but lower yields may result.
3. Separate the Immunomagnetic beads on a Magnetic Separator and save the supernatant containing the target antigen.
4. Repeat Elution step once for higher recovery.
B. Alkaline Elution Protocol
1. Add 100 μL of Alkaline Elution Buffer to the tube.
2. Gently vortex to mix and incubate the sample at room temperature on a rotator for 5 min.
3. Magnetically separate the Immunomagnetic Beads and save the supernatant containing the target antigen.
4. To neutralize the sample, add 50 μL of Neutralization Buffer for each 100 μL of eluate.
C. Acidity Elution Protocol
1. Add 100 μL Acidity Elution Buffer.
2. Gently vortex to mix and incubate the sample at room temperature on a rotator for 5-10 min.
3. Magnetically separate the Immunomagnetic Beads and save the supernatant containing the target antigen.
4. To neutralize the low pH, add 50 μL of Neutralization Buffer for each 100 μL of eluate.
D. Elution Using Sample Buffer
1. Add 100 μL of SDS-PAGE Sample Buffer to the tube.
2. Gently vortex to mix and incubate the sample at 95-100℃ for 5-10 min.
3. Magnetically separate the Immunomagnetic Beads and save the supernatant containing the antigen.
Usage of positive cell lysate
When detecting the antigen with western blotting, the positive cell lysates (supplied in the KIT) can be used as a positive western blotting cell lysates sample.
Note: For tag magnetic beads the positive cell lysates also can be used as a IP cell lysates sample.
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
文献和实验1, Wu L, et al. Fbxo45 facilitates pancreatic carcinoma progression by targeting USP49 for ubiquitination and degradation.Cell death & disease, PubMed ID: 35279684
Cell Sorting Using Immunomagnetic Beads
Immunomagnetic beads are uniform, polymer particles coated with a polystyrene shell that provides both a smooth hydrophobic surface to facilitate physical absorption of molecules, such as antibodies, and surface hydroxyl groups that allow
Dynabeads® Co-Immunoprecipitation Kit
Dynabeads® Co-Immunoprecipitation Kit Magnet: e.g. DynaMag™-2 Mixer allowing rotation or tilting of tubes. Antibodies of your choice. 实验步骤 Day 1 1. Weigh out the appropriate
Sorting of Human Peripheral Blood T-Cell Subsets Using Immunomagnetic Beads
Immunomagnetic beads are uniform, polymer particles coated with a polystyrene shell that provides both a smooth, hydrophobic surface to facilitate absorption of molecules, such as antibodies, and surface hydroxyl groups that allow covalent
技术资料暂无技术资料 索取技术资料
