H-2Dd 抗体
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北京 101111
Quick Overview for H-2Dd 抗体 (ABIN114289)
抗原
适用
宿主
克隆类型
标记
应用范围
克隆位点
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特异性
- This mAb is specific for cells expressing the H-2D antigen coded for by the d haplotype. The reaction pattern of this antibody with a panel of inbred and recombinant haplotypes demonstrates that the antibody detects a private determinant (H-2.4) of the H-2Dd antigen. This antibody can be used to quantitate or eliminate cells bearing H-2Dd (H-2.4) antigen from the appropriate strains of mice.
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纯化方法
- Protein G Chromatography
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免疫原
- Recipient: C3H/HeJ Donor: B6XDBA/2 spleen cells Fusion Partner: SP2/0.Ag14
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亚型
- IgG2a
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anti-MHC Class I H-2Dd antibody
适用: 小鼠 FACS, IP 宿主: 小鼠 Monoclonal 34-5-8S unconjugated
anti-MHC Class I H-2Dd antibody (Biotin)适用: 小鼠 FACS, IP 宿主: 小鼠 Monoclonal 34-5-8S Biotin
anti-MHC Class I H-2Dd antibody (PE)适用: 小鼠 FACS 宿主: 小鼠 Monoclonal 34-5-8S PE
anti-MHC Class I H-2Dd antibody (FITC)适用: 小鼠 FACS 宿主: 小鼠 Monoclonal 34-5-8S FITC
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应用备注
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Cytotoxicity assay. Functional testing.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user. -
实验流程
- RECOMMENDED METHOD FOR DEPLETING A CELL POPULATIONOF H-2Dd BEARING LYMPHOCYTES: 1. Prepare a cell suspension from the appropriate tissue in Cytotoxicity Medium1 orequivalent. Remove red cells and dead cells (where necessary) by purification of viablelymphocytes on Lympholyte®-M density cell separation medium. After washing, adjust thecell concentration to 1x10e7 cells per ml in Cytotoxicity Medium. 2 Add the antibody to afinal concentration of 1: 80 and mix. Alternatively, pellet the cells and resuspend inantibody diluted 1: 80 in Cytotoxicity Medium. 3. Incubate for 60 minutes at 4°C. 4. Centrifuge to pellet the cells and discard the supernatant. 5. Resuspend to the original volume in Low-Tox®-M Rabbit Complement, diluted to theappropriate concentration in Cytotoxicity Medium. (Recommended concentration includedwith each batch of Low-Tox®-M Rabbit Complement. )6. Incubate for 60 minutes at 37°C. 7. Monitor for percent cytotoxicity at this stage, before further processing. For this purposeremove a small sample from each tube, dilute 1: 10 with medium, and add 1/10 volume of1% Trypan Blue. After 3-5 minutes, score live versus dead cells in a haemacytometer. 8. For functional studies, remove the dead cells from the treated groups before furtherprocessing, particularly if the treated cells are to be cultured. This can be done by layeringthe cell suspension separation medium and centrifuging at room temperature as per theinstructions provided. Live cells will form a layer at the interface, while the dead cellspellet. The interface can then be collected and washed in Cytotoxicity Medium before beingresuspended in the appropriate medium for further processing. Alternately, the cells canbe washed and resuspended in the appropriate medium for further processingimmediately after Step 6. , provided that the dead cells will not interfere with subsequentassays. RECOMMENDED METHOD FOR DETERMINING PERCENT OF H-2Dd BEARING CELLS IN A POPULATION: 1. Prepare a cell suspension from the appropriate tissue in Cytotoxicity Medium1 orequivalent. Remove red cells and dead cells (where necessary) by purification of viablelymphocytes on Lympholyte®-M density cell separation medium. After washing, adjust thecell concentration to 1x10e6 cells per ml in Cytotoxicity Medium. 2. Add the antibody to a final concentration of 1: 80 and mix. 3. Incubate for 60 minutes at 4°C. 4. Centrifuge to pellet the cells and discard the supernatant. 5. Resuspend to the original volume in Low-Tox®-M Rabbit Complement3 diluted to theappropriate concentration in Cytotoxicity Medium. (Recommended concentration includedwith each batch of Low-Tox®-M Rabbit Complement. 6. Incubate for 60 minutes at 37°C. 7. Place on ice. 8. Add Trypan Blue, 10% by volume of 1% Trypan Blue (w/v) added 3-5 minutes beforescoring works well. Score live versus dead cells in a haemacytometer. Cytotoxic Index (C. I. )can be calculated as shown in figure 1. NOTES: 1. Cytotoxicity Medium is RPMI-1640 with 25mM Hepes buffer and 0. 3% bovine serumalbumin (BSA). BSA is substituted for the conventionally used fetal calf serum (FCS)because we have found that many batches of FCS contain complement dependentcytotoxins to mouse lymphocytes, thus increasing the background killing in the presenceof complement. We recommend that cells not be exposed to FCS prior to or duringexposure to antibody and complement. Some batches of BSA also contain complement
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限制
- 仅限研究用
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浓度
- 1.0 mg/mL
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缓冲液
- PBS and 0.02 % NaN3
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储存液
- Sodium azide
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注意事项
- This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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注意事项
- Avoid repeated freezing and thawing.
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储存条件
- 4 °C/-20 °C
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储存方法
- Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
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- H-2Dd (MHC Class I H-2Dd)
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别名
- MHC Class I H-2 Dd
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背景
- Synonyms: D-D alpha chain, H-2 class I histocompatibility antigen, H-2D(D), H2-D1
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基因ID
- 100045864
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NCBI登录号
- XP_003086970
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UniProt
- P01900
抗原
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