| Isotype | IgG1 (F)/IgG1(PE) |
| Product Type | Bi-Testª Reagents (FITC/RPE) |
| Units | 100 Tests |
| Host | Mouse |
| Species reactivity | Human |
| Application | Flow Cytometry |
BackgroundIdentification of CD4 on human helper/inducer T cells expressingthe60,000 M.W. surface antigen (HLA class II reactive). CD4 is present in low density on monocytes. Identification of CD8 on human T cells suppressor/cytotoxic expressing the 32,000 M.W. alpha subunit surface antigens.
Synonyms: CD4 FITC - CD8 PE
Source
Immunogen: CD4=Derived from the hybridization of mouse NS-1 myeloma cells with spleen cells from BALB/c mice immunized with human perherial blood T lymphocytes. CD8=Derived from the hybridization of mouse NS-1 myeloma cells with spleen cells from BALB/c mice immunized with human perherial blood T lymphocytes.
Product
Product Form: Bi-Test (FITC/RPE) Reagent
Formulation: Provided as solution in phosphate buffered saline with 0.08% sodium azide and 0.2% carrier protein
Purification Method: Protein A/G Chromatography
Concentration: Titered for flow cytometry
ApplicationsPBMC: Add10 µl of MAB/10^6 PBMC in 100 µl PBS. Mix gently and incubate for 15 minutes at 2º to8ºC. Wash twice with PBS and analyze or fix with 0.5% v/v of paraformaldehyde in PBS and analyze.WHOLE BLOOD: Add10 µl of MAB/100 µl of whole blood.Mix gently and incubate for 15 minutes at room temperature 20ºC. Lyse the whole blood. Wash once with PBS and analyze or fix with 0.5% v/v ofparaformaldehyde in PBS and analyze. See instrument manufacturer’s instructions for Lysed Whole Blood and Immunofluorescence analysis with a flow cytometer or microscope.
Functional Analysis: Flow Cytometry Staining
StorageProduct should be stored at 4-8ºC. DO NOT FREEZE
Product Stability: Reagents are stable for the period shown on the vial label when stored properly
Shipping Conditions: Room Temperature
CautionThis product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Nordic-MUbio accepts no liability for any inaccuracies or omissions in this information.
References1. Thymus Dependent Membrane Antigens in Man: Inhibition of Cell-Mediated Lympholysis by Monoclonal Antibodies to the TH-2 Antigen. Evans, R.L., Wall, D.W., Platsoucas, C.D., Siegal, F.P., Fikrig, S.M., Testa, C.M, and Good, R.A. Proc. Nat. Acad. Sci. 78,544,1981. 2. Novel Immunoregulatory Functions of Phenotypically Distinct Subpopulations of CD4+ cells in the Human Neonate. Clement, L.T., Vink, P.E., Bradley, G.E. J. Immunology 145(1):102-8,1990 . 3. Antigen Presentation by the CD4 Positive Monocyte Subset. Szabo, G.,Miller, C.L., Kodys, K., J. Leukoc. Biol. 47(2): 111-20,1990. 4. Human Immunodeficiency Virus Infection is Efficiently Mediated by a Glycolipid-Anchored form of CD4. Diamond, D.C., Finberg, R., Chaudhuri, S., Sleckman, B.P., Burakoff, S.J., Proc. Natl. Acad. Sci. 87(13):5001-5,1990. 5. Development Regulation of the Intrathymic T cell Precursor Population. Adkins, B.,J. Immunol. 146(5):1387-93,1991. 6. Induction of CD4 and Susceptibility to HIV-1 Infection in Human CD8+ T Lymphocytes by Human Herpesvirus 6. Lusso, P., De Maria, A., Malnati, M., Lori, F., DeRocco, S.E., Baseler, M., Gallo, R.C., Nat. 349(6309):533-5,1991. 7. Evolutionary Conservation of Surface Molecules that Distinguish T Lymphocyte Helper/Inducer and T Cytotoxic/Suppressor Subpopulations in Mouse and Man. Ledbetter,J.A., Evans, R.L., Lipinski,M., Cunningham-Rundles,C., Good, R.A., and Herzenberg, L.A., J. Exp. Med. 153,310, 1981. 8. Circulating Antigen-Specific Suppressor T Cells in a Healthy Woman: Mechanism of Action and Isolation with a Monoclonal Antibody. Engleman, E.G., Benike, C.J., and Evans, R.L., Clin. Res. 29, 365a 1981. 9. Induction of Immunoglobulin Secreting Cells in the Allogeneic Mixed Leukocyte Reaction: Regulation by Helper and Suppressor Lymphocyte Subsets in Man. Kotzin, B.L., Benike, C.J. and Engleman, E.G., J. Imm. 127,931,1981. 10. CD4 and CD8 molecules can physically associate with the same T-cell receptor. Gallagher, P.F., Fazekas de St. Groth, B.,Miller,J.F.A.P., Proc. Nat. Acad. Sci. 1989,86:10044.
Protein Reference(s)
Database Name: UniProt
Accession number: P01730, P01732
Safety Datasheet(s) for this product:NM_Sodium Azide/wp-content/uploads/SDS/Antibody SDS with Sodium Azide Noridic-MUbio.pdf
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其次,察看次目的蛋白的存在形式,有没有多聚体形式及变构形式;
最后,查看多家抗体公司的DATA,看看别人的WB做出来的条带的位置。
根据你说的,特异识别多个组织中的同样大小的条带,我觉得很可能就是你的目的蛋白。
一般来说多克隆的阳性率高一些,但出现假阳性的比例也高一些。
1取组织
(1) 灌注固定:暴露心脏,将注射针头插入左心室,剪开右心耳,快速灌注PBS,待右心耳处流出澄清液体,换4%多聚甲醛灌注到大鼠四肢强直为止,快速取脑和肾。
(2) 固定:将组织置于4%多聚甲醛中固定15小时
(3) 脱水:将组织放入30% 蔗糖溶液中直至下沉
(4) 切片:脑组织冰冻切片20微米,37度烘干2h以上;
(5)将切片放置-80度保存,随用随取。
2免疫单标记
(1)做抗原修复(0.4g柠檬酸、3g柠檬酸钠、1000ml水配制成溶液,放到水浴箱中90~94℃保存15min),冷却后, 用1*PBS反复洗标本10min,吸去PBS,同时向标本加渗透液(配0.4%trition液,然后将BSA加入到配好的trition液中,配成1%浓度)100ul,常温下湿盒反应1h。
(2) 吸去渗透液,同时向标本加block液(5%封闭用正常山羊血清)100ul,常温下湿盒反应5h。
(3) 吸去block液,同时向标本加入1:50稀释的羊抗鼠一抗100ul,37度湿盒反应过夜。
(4)吸去一抗,并回收,用1*PBS洗液反复洗15min。
(5)快速从冰箱中取出驴抗羊的二抗,并立即用锡纸包裹,注入到一个新的以锡纸包裹的tube管中并用PBS以1:200稀释。
(6)进入暗室,吸去PBS,同时加入二抗100ul,37度湿盒反应2h,封片。
(7)放置到荧光显微镜下观察。
又由于自然存在的抗原大都存在多个抗原表位,会刺激机体产生多种针对同一抗原的不同抗原表位相应的不同抗体.

