Re: [Histonet] shelf life of working antibody solutions in IHC
I run a research lab with automated staining and truthfully,I make my dilutions fresh for each run because proteins are so different in their stability. Colleen Forster HT(ASCP)QIHC On Sun, Jul 2, 2023 at 7:08 PM Tony Henwood via Histonet < histonet@lists.utsouthwestern.edu> wrote: > In Australia, we have evolved from Prescriptive accreditation (following > the standards) to Risk-based accreditation where it is more important to > assess the risk of a laboratory process rather than only meeting a list of > rules, since following a list of rules is often not in the patient’s nor in > the community’s interest. > > Years ago, I battled with our accreditation organization over the risk > involved in fridge temperature recording, presenting evidence that > continuous digital monitoring (with an alarm system) was less risky than > recording max/min temperatures daily (eg what happens when the lab is not > manned during Christmas holidays and weekends?). > > The excessive cost involved in automatically disposing of expired > antibodies is wasteful. Continuous monitoring as well as preventative > practices such as preservatives (azide, procillin, etc) and pH monitoring > with phenol red greatly reduces the risk. > > As presented in the paper, many working dilutions survive without > contamination for many years. For example, factor 8 (21 years), factor 13a > (19 years) and epithelial membrane antigen (17 years). There were no failed > verifications and to date, the average life after expiration was 6 years; > eight antibodies exceeded 6 years. All the other antibodies would probably > have similar survival times but were exhausted before reaching these times. > > Regards, > > Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired) > Principal Scientist, the Children’s Hospital at Westmead (Retired) > Adjunct Fellow, School of Medicine, University of Western Sydney. > > From: Gudrun Lang<mailto:gu.l...@gmx.at> > Sent: Sunday, 2 July 2023 7:26 PM > To: 'Tony Henwood'<mailto:afhenw...@outlook.com> > Subject: AW: [Histonet] shelf life of working antibody solutions in IHC > > Dear Tony, > thank you for the interesting article. It reflects my personal experience. > Nevertheless accreditation rules overrule the experience and in an > diagnostic lab we have to discard the reagenses. > > My question regards more the practical approach with working solutions. > What is your experience how long the working solutions are „good enough“ > for using. > > Thank you very much and kind regards > Gudrun > > Von: Tony Henwood [mailto:afhenw...@outlook.com] > Gesendet: Samstag, 1. Juli 2023 23:41 > An: Gudrun Lang > Betreff: RE: [Histonet] shelf life of working antibody solutions in IHC > > Here it is > > Regards, > > Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired) > Principal Scientist, the Children’s Hospital at Westmead (Retired) > Adjunct Fellow, School of Medicine, University of Western Sydney. > > From: Gudrun Lang via Histonet<mailto:histonet@lists.utsouthwestern.edu> > Sent: Sunday, 2 July 2023 4:59 AM > To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu> > Subject: [Histonet] shelf life of working antibody solutions in IHC > > Hi all! > > I have tried to find a general instruction for the shelf life of antibody > working solutions. > > With automated IHC you usually fill the container with the working solution > and depending on the frequency of usage they stay on the instrument at > roomtemperature (or higher) or are put in the fridge again. > > The working solutions are up to 10 ml and may last for months. The > antibody-diluent is from the same company of the instrument. The titers are > in a range from 1:10 to 1:3000. - so a very heterogen situation. > > > > How do you handle this? Have you a general rule, when the solution has to > be > discarded? Is it just a matter of positiv-controls? > > > > Thanks in advance > > Gudrun Lang > > Biomedical scientist > > Austria > > > > > > ___ > 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 > -- Colleen Forster HT(ASCP)QIHC BLS Histology and IHC Laboratory Jackson Hall, Room 2-155 321 Church St. SE Minneapolis, MN 55455 612-626-1930 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] shelf life of working antibody solutions in IHC
In Australia, we have evolved from Prescriptive accreditation (following the standards) to Risk-based accreditation where it is more important to assess the risk of a laboratory process rather than only meeting a list of rules, since following a list of rules is often not in the patient’s nor in the community’s interest. Years ago, I battled with our accreditation organization over the risk involved in fridge temperature recording, presenting evidence that continuous digital monitoring (with an alarm system) was less risky than recording max/min temperatures daily (eg what happens when the lab is not manned during Christmas holidays and weekends?). The excessive cost involved in automatically disposing of expired antibodies is wasteful. Continuous monitoring as well as preventative practices such as preservatives (azide, procillin, etc) and pH monitoring with phenol red greatly reduces the risk. As presented in the paper, many working dilutions survive without contamination for many years. For example, factor 8 (21 years), factor 13a (19 years) and epithelial membrane antigen (17 years). There were no failed verifications and to date, the average life after expiration was 6 years; eight antibodies exceeded 6 years. All the other antibodies would probably have similar survival times but were exhausted before reaching these times. Regards, Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired) Principal Scientist, the Children’s Hospital at Westmead (Retired) Adjunct Fellow, School of Medicine, University of Western Sydney. From: Gudrun Lang<mailto:gu.l...@gmx.at> Sent: Sunday, 2 July 2023 7:26 PM To: 'Tony Henwood'<mailto:afhenw...@outlook.com> Subject: AW: [Histonet] shelf life of working antibody solutions in IHC Dear Tony, thank you for the interesting article. It reflects my personal experience. Nevertheless accreditation rules overrule the experience and in an diagnostic lab we have to discard the reagenses. My question regards more the practical approach with working solutions. What is your experience how long the working solutions are „good enough“ for using. Thank you very much and kind regards Gudrun Von: Tony Henwood [mailto:afhenw...@outlook.com] Gesendet: Samstag, 1. Juli 2023 23:41 An: Gudrun Lang Betreff: RE: [Histonet] shelf life of working antibody solutions in IHC Here it is Regards, Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired) Principal Scientist, the Children’s Hospital at Westmead (Retired) Adjunct Fellow, School of Medicine, University of Western Sydney. From: Gudrun Lang via Histonet<mailto:histonet@lists.utsouthwestern.edu> Sent: Sunday, 2 July 2023 4:59 AM To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: [Histonet] shelf life of working antibody solutions in IHC Hi all! I have tried to find a general instruction for the shelf life of antibody working solutions. With automated IHC you usually fill the container with the working solution and depending on the frequency of usage they stay on the instrument at roomtemperature (or higher) or are put in the fridge again. The working solutions are up to 10 ml and may last for months. The antibody-diluent is from the same company of the instrument. The titers are in a range from 1:10 to 1:3000. - so a very heterogen situation. How do you handle this? Have you a general rule, when the solution has to be discarded? Is it just a matter of positiv-controls? Thanks in advance Gudrun Lang Biomedical scientist Austria ___ 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] shelf life of working antibody solutions in IHC
Hi Tony, Very interesting article you shared. Hi Gudrun, I will only add to this - run positive and negative tests for every run. Good luck! Naira On Sat, Jul 1, 2023 at 4:40 PM Tony Henwood via Histonet < histonet@lists.utsouthwestern.edu> wrote: > Hi Gudrun, > > This should be useful ( I will send a copy separately) > > Henwood, A. F. (2023). Validation of nominally expired antibodies for > immunohistochemistry. Biotechnic & Histochemistry, 98(2), 86-93. > > Regards, > > Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired) > Principal Scientist, the Children’s Hospital at Westmead (Retired) > Adjunct Fellow, School of Medicine, University of Western Sydney. > > From: Gudrun Lang via Histonet<mailto:histonet@lists.utsouthwestern.edu> > Sent: Sunday, 2 July 2023 4:59 AM > To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu> > Subject: [Histonet] shelf life of working antibody solutions in IHC > > Hi all! > > I have tried to find a general instruction for the shelf life of antibody > working solutions. > > With automated IHC you usually fill the container with the working solution > and depending on the frequency of usage they stay on the instrument at > roomtemperature (or higher) or are put in the fridge again. > > The working solutions are up to 10 ml and may last for months. The > antibody-diluent is from the same company of the instrument. The titers are > in a range from 1:10 to 1:3000. - so a very heterogen situation. > > > > How do you handle this? Have you a general rule, when the solution has to > be > discarded? Is it just a matter of positiv-controls? > > > > Thanks in advance > > Gudrun Lang > > Biomedical scientist > > Austria > > > > > > ___ > 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 > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] shelf life of working antibody solutions in IHC
Hi Gudrun, This should be useful ( I will send a copy separately) Henwood, A. F. (2023). Validation of nominally expired antibodies for immunohistochemistry. Biotechnic & Histochemistry, 98(2), 86-93. Regards, Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired) Principal Scientist, the Children’s Hospital at Westmead (Retired) Adjunct Fellow, School of Medicine, University of Western Sydney. From: Gudrun Lang via Histonet<mailto:histonet@lists.utsouthwestern.edu> Sent: Sunday, 2 July 2023 4:59 AM To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: [Histonet] shelf life of working antibody solutions in IHC Hi all! I have tried to find a general instruction for the shelf life of antibody working solutions. With automated IHC you usually fill the container with the working solution and depending on the frequency of usage they stay on the instrument at roomtemperature (or higher) or are put in the fridge again. The working solutions are up to 10 ml and may last for months. The antibody-diluent is from the same company of the instrument. The titers are in a range from 1:10 to 1:3000. - so a very heterogen situation. How do you handle this? Have you a general rule, when the solution has to be discarded? Is it just a matter of positiv-controls? Thanks in advance Gudrun Lang Biomedical scientist Austria ___ 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
[Histonet] shelf life of working antibody solutions in IHC
Hi all! I have tried to find a general instruction for the shelf life of antibody working solutions. With automated IHC you usually fill the container with the working solution and depending on the frequency of usage they stay on the instrument at roomtemperature (or higher) or are put in the fridge again. The working solutions are up to 10 ml and may last for months. The antibody-diluent is from the same company of the instrument. The titers are in a range from 1:10 to 1:3000. - so a very heterogen situation. How do you handle this? Have you a general rule, when the solution has to be discarded? Is it just a matter of positiv-controls? Thanks in advance Gudrun Lang Biomedical scientist Austria ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet