[Histonet] How to save dried antibody?
Dear All, We have purchased a sheep anti-CGRP antibody for our fluorescent IHC study on human tissue and kept it in -20 for a while. It came in liquid but now it's all stiff and looks like it's being crystallized. I cannot pipet any of it. It has hardened and looks like a piece of glass at bottom of its container. I tried to re-suspend it with the same amount of glycerol, but the antibody doesn't seem to be mixing with it. Does anyone know how to save this antibody? How can I bring it back to its liquid state? Thank you for your time Sarah Sadat, DDS MSc Student, Dental Sciences McGill University Montreal QC. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] How to save dried antibody?
Not sure if others would call this a lyophilized situation. Dont know the specifics of what you initially recieved. Here's my best shot as I recall from using lyophilized antibodies doing IF for renal bx's a while back. Back then I used a PBS based diluent. A very low reconstitution amount, 50-100um. I typically used the diluent in the pipett tip to swirl the freeze dried/lyophilized antibody back together while warming the tube with my thumb as I held it. It worked well in that situation for me. Hope this helps. I'm sure there are others out here who do this every day and can offer more specifics? Best of luck. Kim Donadio From: sarah Tabatabaei sarah_t...@yahoo.com To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Friday, October 28, 2011 11:27 AM Subject: [Histonet] How to save dried antibody? Dear All, We have purchased a sheep anti-CGRP antibody for our fluorescent IHC study on human tissue and kept it in -20 for a while. It came in liquid but now it's all stiff and looks like it's being crystallized. I cannot pipet any of it. It has hardened and looks like a piece of glass at bottom of its container. I tried to re-suspend it with the same amount of glycerol, but the antibody doesn't seem to be mixing with it. Does anyone know how to save this antibody? How can I bring it back to its liquid state? Thank you for your time Sarah Sadat, DDS MSc Student, Dental Sciences McGill University Montreal QC. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] How to save dried antibody?
If the antibody has precipitated, it may be impossible to restore activity. The glycerol is present to keep the suspension in liquid form - and it sounds like you no longer have a liquid. Try adding an appropriate suspension buffer and keep your fingers crossed. If you do not have previous experience with this antibody and know what to expect regarding performance, I would discard it and re-order. Tresa -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sarah Tabatabaei Sent: Friday, October 28, 2011 9:28 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] How to save dried antibody? Dear All, We have purchased a sheep anti-CGRP antibody for our fluorescent IHC study on human tissue and kept it in -20 for a while. It came in liquid but now it's all stiff and looks like it's being crystallized. I cannot pipet any of it. It has hardened and looks like a piece of glass at bottom of its container. I tried to re-suspend it with the same amount of glycerol, but the antibody doesn't seem to be mixing with it. Does anyone know how to save this antibody? How can I bring it back to its liquid state? Thank you for your time Sarah Sadat, DDS MSc Student, Dental Sciences McGill University Montreal QC. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] How to save dried antibody?
I personally would not use it; you don't know how much loss of immunologic reactivity has occurred, and that might affect your project data. Buy a fresh supply, if possible. Jan Shivers Senior Scientist Histology/IHC/EM Section Head Pathology Teaching Program University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive...@umn.edu (Confidentiality Notice: This message, together with any attachments, is intended only for the use of the individual or entity to which it is addressed and may contain confidential or privileged information. If you think you have received this message in error, please advise the sender and then delete this message and any attachments immediately.) - Original Message - From: sarah Tabatabaei sarah_t...@yahoo.com To: histonet@lists.utsouthwestern.edu Sent: Friday, October 28, 2011 10:27 AM Subject: [Histonet] How to save dried antibody? Dear All, We have purchased a sheep anti-CGRP antibody for our fluorescent IHC study on human tissue and kept it in -20 for a while. It came in liquid but now it's all stiff and looks like it's being crystallized. I cannot pipet any of it. It has hardened and looks like a piece of glass at bottom of its container. I tried to re-suspend it with the same amount of glycerol, but the antibody doesn't seem to be mixing with it. Does anyone know how to save this antibody? How can I bring it back to its liquid state? Thank you for your time Sarah Sadat, DDS MSc Student, Dental Sciences McGill University Montreal QC. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
