From hqian from mail.mcg.edu Tue Sep 2 09:34:03 2008 From: hqian from mail.mcg.edu (Hailun Qian) Date: Tue Sep 2 10:55:52 2008 Subject: [Cell-biology] bcl2 expressing jurket cell line&In-Reply-To= Message-ID: <48BD174E.EFEC.0025.0@mail.mcg.edu> Dear Sir/madam: Please tell me where I can get Bcl-2 expressing jurket cells ? Helen From a.meedeniya from griffith.edu.au Thu Sep 4 19:48:37 2008 From: a.meedeniya from griffith.edu.au (Adrian Meedeniya) Date: Fri Sep 5 10:43:04 2008 Subject: [Cell-biology] HOXB6 Antibody Message-ID: Dear Dr Anderson, We have encountered difficulties in immunostaining for HOXB6 in mouse and human stem cells and would be very grateful for access to some of the antibody generated by your group. Could you please provide information on costs and logistics, for supply of the antibody.. Thank you, with best regards, Adrian __________________________________ Dr Adrian C. B. Meedeniya National Centre for Adult Stem Cell Research Griffith University Eskitis Building N75 Brisbane Innovation Park, Don Young Road, Nathan, Queensland, 4111, Australia ph: +61 7 3735 4452 (Lab) / 3735 4417 (Office) Fax: +61 7 3735 4255 email: A.Meedeniya@griffith.edu.au From kcarbaja from uci.edu Tue Sep 16 17:08:05 2008 From: kcarbaja from uci.edu (Kevin Carbajal) Date: Tue Sep 16 17:25:23 2008 Subject: [Cell-biology] IHC CD4 and CD8 staining Message-ID: <8bc739800809161508y6c899fc1ub28a3e38be2726c7@mail.gmail.com> Hello,I'm trying to stain for CD4 and CD8 antigens in mouse spinal cords (that have vast T-cell infiltration) and can't get it to work. My tissue is frozen and I have used it successfully in staining many other antigens, but these don't show. My primaries are rat anti-mouse antibodies from Pharmingen (for IHC) and I have used two secondaries: a donkey anti-rat PE, and a biotinylated rabbit anti-rat. I've fixed the tissues with 4% Paraformaldehyde for 20 min, and with acetone at -20C for 10 min. Blocked with 5-10% of the corresponding serum and followed standard protocols (washing, mounting, DAB staining in the case of the biotin. ab, etc). My incubations are over-night in 4C for the primaries and 1.5 hrs RT for the secondaries. I've used diverse concentrations of the antibodies. I have also tried it all on spleen sections for positive controls. What could I be doing wrong? Do I need to buy new antibodies? Thank you for any suggestions! -- Kevin S. Carbajal From ashishviro from gmail.com Tue Sep 23 10:31:23 2008 From: ashishviro from gmail.com (ashish tiwari) Date: Tue Sep 23 11:31:44 2008 Subject: [Cell-biology] antibody response to lysozyme Message-ID: <93ea52690809230831h1f33ac0fi969b081860bc933d@mail.gmail.com> hi, please help me to understand this HyHEL-10 hybridoma produces a monoclonal antibody that is specific for hen egg lysozyme. HyHEL-10 antibody binds to hen egg lysozyme (HEL) with an affinity of KA~4.5x1010 M-1, duck egg lysozyme (DEL) with an affinity of KA ~1x107M-1. Transgenic mice called MD4 express HyHEL-10 as its B cell receptor. Answer the following questions. a. What would you predict is the difference between HEL and DEL and how would this affect affinity to HyHEL-10? b. Draw a graph with antibody levels on the y-axis and time on the x-axis showing the relative antibody responses to HEL and DEL in MD4 transgenic mice. Explain the reasoning behind your graph. c. How would the graph in b. change if you used MD4 mice crossed to CR2 knockout mice? -- Ashish Tiwari