MHC Class I (RT1Ac) 抗体
Quick Overview for MHC Class I (RT1Ac) 抗体 (ABIN114395)
抗原
适用
宿主
克隆类型
标记
应用范围
克隆位点
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纯化方法
- Protein G Chromatography
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免疫原
- Phytohaemagglutinin Blasts Donor: BALB/c spleen Fusion Partner: P3-NS1-1-Ag4 (NS1/1)
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亚型
- IgG2a
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应用备注
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Flow Cytometry. Immunoprecipitation. Immunohistochemistry using cryostat sections: (however, cross-reactivity with Lewis ratshave been shown to occur in some instances).
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 a 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 the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test). 4. To each tube, add 0. 5-2. 0 µg 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: 500 dilution. Notrecommended for use with PE secondary. 9. Incubate the 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 PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results - Tissue Distribution: Rat Strain: Brown NorwayCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 5 µg/10e6 cellsIsotypic Control: Mouse IgG2aCell Source Percentage of cells stained above control: Thymus 39. 8%Spleen 95. 9%Lymph Node 99. 9%Results - Strain Distribution: Cell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 5 µg/10e6 cellsStrains Tested: Lewis, Wistar, Brown Norway, Fischer 344, Buffalo, ACIPositive: Brown NorwayNegative: Lewis, Wistar, Fischer 344 Buffalo, ACI
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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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- MHC Class I (RT1Ac)
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背景
- MHC Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. MHC class I antigens are heterodimers consisting of one alpha chain (44 kDa) with beta 2 microglobulin (11.5 kDa). The antigen is expressed by all somatic cells at varying levels. MHC Class I molecules are expressed on most nucleated cells where they present endogenously synthesized antigenic peptides to CD8+ T lymphocytes, which are usually cytotoxic T cells. Fibroblasts or neurons however only show a low level of antigen.Synonyms: MHC class I RT1.Ac heavy chain, RT1-A3
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基因ID
- 10116
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UniProt
- Q31255
抗原
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