(5 references)-
- 抗原 See all CD4 products
- CD4
- 适用
- 小鼠
- 宿主
- 大鼠
- 克隆类型
- 单克隆
- 标记
- Magnetic Particles
- 应用范围
- Separation (Sep)
- 品牌
- BD IMag™
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- 说明
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Blocking, Depletion, Immunoprecipitation
- 实验流程
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1. Prepare a single-cell suspension from the lymphoid tissue of interest according to standard laboratory procedures. Remove clumps of cells and/or debris by passing the suspension through a 70-µm nylon cell strainer.
2. Dilute BD IMag™ Buffer (10X) (Cat. No. 552362) 1:10 with sterile distilled water or prepare 1X BD IMag™ buffer by supplementing Phosphate Buffered Saline with 0.5% BSA, 2 mM EDTA, and 0.09% sodium azide. Place on ice. Although our experience indicates that the use of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141) is not required for optimal cell separation, some laboratories may want to use it in their studies. If adding Mouse BD Fc Block™, proceed to Step 3. If not adding Mouse BD Fc Block™, proceed to Step 4.
3. Add Mouse BD Fc Block™ at 0.25 µg/10e6 cells, and incubate on ice for 15 minutes.
4. Wash cells with at least an equal volume of 1X BD IMag™ buffer, and carefully aspirate all the supernatant.
5. Vortex the BD IMag™ anti-mouse CD4 Magnetic Particles - DM thoroughly, and add 50 µl of particles for every 10e7 total cells.
6. MIX THOROUGHLY. Refrigerate at 6°C - 12°C for 30 minutes.
7. Bring the BD IMag™-particle labeling volume up to 1 - 8 x 10e7 cells/ml with 1X BD IMag™ buffer, and immediately place the tube on the BD IMagnet™. Incubate at room temperature for 6 - 8 minutes.
8. With the tube on the BD IMagnet™, carefully aspirate off the supernatant. This supernatant contains the negative fraction.
9. Remove the tube from the BD IMagnet™, and add 1X BD IMag™ buffer to the same volume as in Step 7. Gently resuspend cells by pipetting briefly, and return the tube to the BD IMagnet™ for another 2 - 4 minutes.
10. With the tube on the BD IMagnet™, carefully aspirate off the supernatant and discard.
11. Repeat Steps 9 and 10.
12. After the final wash step, resuspend the positive fraction in an appropriate buffer and at an appropriate concentration for further analysis.
NOTE: Avoid nonspecific labeling by working quickly and adhering to recommended incubation times. - 限制
- 仅限研究用
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- 状态
- Liquid
- 缓冲液
- Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.
- 储存液
- Sodium azide
- 注意事项
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- 储存条件
- 4 °C
- 储存方法
- Store undiluted at 4°C.
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: "Increased regulatory versus effector T cell development is associated with thymus atrophy in mouse models of multiple myeloma." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 181, Issue 5, pp. 3714-24, (2008) (PubMed).
: "Mouse NK1+ T cells." in: Current opinion in immunology, Vol. 7, Issue 3, pp. 367-74, (1995) (PubMed).
: "CD4 is expressed on murine pluripotent hematopoietic stem cells." in: Blood, Vol. 80, Issue 7, pp. 1717-24, (1992) (PubMed).
: "L3T4 antigen expression by hemopoietic precursor cells." in: The Journal of experimental medicine, Vol. 169, Issue 4, pp. 1473-8, (1989) (PubMed).
: "Characterization of the murine T cell surface molecule, designated L3T4, identified by monoclonal antibody GK1.5: similarity of L3T4 to the human Leu-3/T4 molecule." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 131, Issue 5, pp. 2445-51, (1983) (PubMed).
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: "Increased regulatory versus effector T cell development is associated with thymus atrophy in mouse models of multiple myeloma." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 181, Issue 5, pp. 3714-24, (2008) (PubMed).
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- 抗原
- CD4
- 别名
- CD4 (CD4 产品)
- 别名
- cd4 Accessory Reagents, L3T4 Accessory Reagents, Ly-4 Accessory Reagents, CD4mut Accessory Reagents, W3/25 Accessory Reagents, p55 Accessory Reagents, CD4 molecule Accessory Reagents, carcinoembryonic antigen-related cell adhesion molecule 5 Accessory Reagents, T-cell surface glycoprotein CD4 Accessory Reagents, CD4 antigen Accessory Reagents, Cd4 molecule Accessory Reagents, CD4 Accessory Reagents, LOC100125525 Accessory Reagents, cd4 Accessory Reagents, Cd4 Accessory Reagents
- 背景
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BD IMag™ anti-mouse CD4 Particles - DM are magnetic nanoparticles that have monoclonal antibody conjugated to their surfaces. These particles are optimized for the positive selection or depletion of CD4-bearing leukocytes using the BD IMagnet™. The CD4 (L3T4) differentiation antigen has been reported to be expressed on most thymocytes, a subpopulation of mature T lymphocytes (i.e., MHC class II-restricted T cells, including most T helper cells), and a subset of NK-T cells. In addition, CD4 has also been reported to be detectable on pluripotent hematopoietic stem cells, bone marrow myeloid and B-lymphocyte precursors, intrathymic lymphoid precursors, and a subset of splenic dendritic cells. Leukocytes are labeled with BD IMag™ anti-mouse CD4 Particles - DM according to the Magnetic Labeling Protocol. This labeled cell suspension is then placed within the magnetic field of the BD IMagnet™ (Cat.No. 552311). Labeled cells migrate toward the magnet (positive fraction), leaving the unlabeled cells in suspension so they can be drawn off (negative fraction). The tube is then removed from the magnetic field for resuspension of the positive fraction. The separation is repeated twice to increase the purity of the positive fraction. The magnetic separation steps are diagrammed in the Separation Flow Chart. After the positive fraction is washed, the small size of the magnetic particles allows the positive fraction to be further evaluated in downstream applications such as flow cytometry.
Synonyms: L3T4
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