MHC Class I H-2Dd 抗体

MHC Class I H-2Dd 抗体抗原
MHC Class I H-2Dd
适用
其他选择 小鼠
宿主
其他选择 小鼠
克隆类型 (克隆位点)
单克隆 (34-5-8S)
标记
其他选择 非结合性
应用范围
其他选择 Cytotoxicity Test (CyTox)
Pubmed 找到1个引用
规格 0.25 mg
发货至 中国 (更改)
发货时间 29至35个工作日
产品编号 yb114289
产品细节 MHC Class I H-2Dd 抗体
免疫原 Recipient: C3H/HeJ Donor: B6XDBA/2 spleen cells Fusion Partner: SP2/0.Ag14
克隆位点 34-5-8S
亚型 IgG2a
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.
纯化方法 Protein G Chromatography
目标详细情况 MHC Class I H-2Dd 抗体
别名 MHC Class I H-2 Dd
背景 Synonyms: D-D alpha chain, H-2 class I histocompatibility antigen, H-2D(D), H2-D1
基因ID 100045864
NCBI登录号 XP_003086970
UniProt P01900
使用细节 MHC Class I H-2Dd 抗体
应用备注 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
限制 仅限研究用
贮存及处理 MHC Class I H-2Dd 抗体
浓度 1.0 mg/mL
缓冲液 PBS and 0.02 % NaN3
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.
注意事项 Avoid repeated freezing and thawing.
储存条件 4 °C/-20 °C
储存方法 Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.