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- 抗原
- MHC Class II (RT1D)
- 适用
- 大鼠
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宿主
- 小鼠
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克隆类型
- 单克隆
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标记
- 非结合性
- 应用范围
- Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunoprecipitation (IP)
- 纯化方法
- Protein G affinity chromatography
- 免疫原
- Rat spleen membrane glycoproteins depleted of Ia-A antigens Immunocyte Donor: BALB/c spleen Fusion Partner: X63 Ag8.653
- 克隆位点
- OX-17
- 亚型
- IgG1
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anti-MHC Class II (RT1D) antibody (Biotin)
适用: 大鼠 FACS, IHC (fro) 宿主: 小鼠 Monoclonal OX-17 Biotin
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- 应用备注
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Flow Cytometry: 1/250 - 1/500 (see Protocols). Immunhistochemistry. Immunoprecipitation.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user. - 实验流程
- FLOW CYTOMETRY ANALYSIS: METHOD: 1. Prepare cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-Rat. cell separation medium. 2. Wash 2 times. 3. Resuspend cells to 1x10e6 cells in approximately 50 µl media A in a microcentrifugetube. (i. e. 50 µl of cells resuspended to 2x10e7 cells/ml). The contents of 1 tube represent 1test. 4. To each tube add 50 µl of a 1: 250 - 1: 500 dilution of this Ab. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (FITC Goat anti-mouse IgG (H+L)) at 1: 700 dilution. 9. Incubate tubes at 4°C for 30-60 minutes. (It is recommended that the tubes areprotected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in Media B. 11. Resuspend the cell pellet in 50 µl ice cold Media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in phosphate buffered saline. (This stains dead cells by intercalatingDNA). MEDIA: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2 M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% bovine serum albumin + sodium azide (100 µlof 2 M sodium azide in 100 mls). RESULTS: Rat Strain: Lewis RatCell Concentration: 1x10e6 cells per testAntibody Concentration: 1: 400Isotypic Control: Mouse IgG1, κCELL SOURCE PERCENT STAININGThymus 12%Spleen 34%Lymph Node 25%STRAIN DISTRIBUTION: Antibody Concentration: 1: 100Strains Tested: Wistar, Buffalo, Brown Norway, Fischer 344Positive: Wistar, Buffalo, Brown Norway, ACI, Fischer 344Negative: none
- 限制
- 仅限研究用
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- 浓度
- 1.0 mg/mL
- 缓冲液
- PBS with 0.02 % 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.
- 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.
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- 抗原
- MHC Class II (RT1D)
- 别名
- MHC Class II RT1D
- 背景
- Synonyms: HLA Class II
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