(4 references)-
- 抗原 See all CD11b (ITGAM) products
- CD11b (ITGAM) (Integrin alpha M (ITGAM))
- 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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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 CD11b 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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: "Flow cytometric identification of murine neutrophils and monocytes." in: Journal of immunological methods, Vol. 197, Issue 1-2, pp. 139-50, (1996) (PubMed).
: "Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors." in: Science (New York, N.Y.), Vol. 245, Issue 4923, pp. 1238-41, (1989) (PubMed).
: "Cross-reaction of a rat-anti-mouse phagocyte-specific monoclonal antibody (anti-Mac-1) with human monocytes and natural killer cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 126, Issue 1, pp. 359-64, (1981) (PubMed).
: "Mac-1: a macrophage differentiation antigen identified by monoclonal antibody." in: European journal of immunology, Vol. 9, Issue 4, pp. 301-6, (1979) (PubMed).
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: "Flow cytometric identification of murine neutrophils and monocytes." in: Journal of immunological methods, Vol. 197, Issue 1-2, pp. 139-50, (1996) (PubMed).
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- 抗原
- CD11b (ITGAM) (Integrin alpha M (ITGAM))
- 别名
- CD11b (ITGAM 产品)
- 别名
- CD11b/CD18 Accessory Reagents, CR3 Accessory Reagents, CR3A Accessory Reagents, Cd11b Accessory Reagents, F730045J24Rik Accessory Reagents, Ly-40 Accessory Reagents, MAC1 Accessory Reagents, Mac-1 Accessory Reagents, Mac-1a Accessory Reagents, CD11B Accessory Reagents, MAC-1 Accessory Reagents, MAC1A Accessory Reagents, MO1A Accessory Reagents, SLEB6 Accessory Reagents, C3bi Accessory Reagents, integrin subunit alpha M Accessory Reagents, integrin alpha M Accessory Reagents, integrin, alpha M (complement component 3 receptor 3 subunit) Accessory Reagents, integrin alpha-M Accessory Reagents, ITGAM Accessory Reagents, Itgam Accessory Reagents, LOC100351865 Accessory Reagents
- 背景
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BD IMag™ anti-mouse CD11b Particles - DM are magnetic nanoparticles that have monoclonal antibody conjugated to their surfaces. These particles are optimized for the positive selection or depletion of CD11b-bearing leukocytes using the BD IMagnet™. CD11b is expressed at varying levels on granulocytes, macrophages, myeloid-derived dendritic cells, natural killer cells, and B-1 cells and is rapidly upregulated on neutrophils after activation. Leukocytes are labeled with BD IMag™ anti-mouse CD11b 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: Integrin alpha[M] chain, Mac-1 alpha chain, CR3
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