6 replies to this topic

[human]

I'm troblemed in immunohistchemistry of mouse monoclonal antibody for beta-galactosidase produced by Promega. I tryed staining of mouse tissue twice to both frosen section and paraffin section using M.O.M kit. 1:300 and 1:2000 dilution, I tried. But no signal was detected in paraffin section and high background signal in frozen section. If anyone can succeed in the immunohistochemistry using this antibody, please tell me how to stain in detail or troubleshooting.

[dako]

Hi, I have same problem, please write me dilution of this antibody from promega. Before I used beta gal. from Chemicon, it was rabbit dilution 1:500 and from Sanes (rabbit) 1:800. It was working good, but better in brightfiedl. Thanks for answer, bye

[human]

dako, on Nov 4 2004, 08:30 AM, said:

Hi, I have same problem, please write me dilution of this antibody from promega. Before I used beta gal. from Chemicon, it was rabbit dilution 1:500 and from Sanes (rabbit) 1:800. It was working good, but better in brightfiedl. Thanks for answer, bye

Thank you for your valuable imformation. After I hadn't succeed in the antibody of Promega, I also tried Chemicon's one. One previous report had described the appropriate dilution was 1:1500, but I couldn't get good result. Next time I'm going to try 1:500. Good bye.

[dako]

now, I am trying beta gal. Promega 1:200, 1:1000, 1:2000 I use just 1 reagent from M.O.M kit - Blocking reagent, preincubation with Normal Goat Serum and secondary antibody I take Goat anti Mouse Peroxidase (Sigma), so I am curiouse, When I will have result I will write you. Where are zou come from? And which language do you speak? Bye

[human]

dako, on Nov 5 2004, 09:57 AM, said:

now, I am trying beta gal. Promega 1:200, 1:1000, 1:2000 I use just 1 reagent from M.O.M kit - Blocking reagent, preincubation with Normal Goat Serum and secondary antibody I take Goat anti Mouse Peroxidase (Sigma), so I am curiouse, When I will have result I will write you. Where are zou come from? And which language do you speak? Bye

For using the Promega antibody, I also used M.O.M. kit, including blocking reagent and working solution. After that I chose ABC kit for secondary antibody of anti-mouse peroxidase. But the difference is very small. Please tell me your result. Good luck!
I am from Japan, I'm a second grade graduate school student at the department of Genetics. Good bye.

[dako]

human, on Nov 6 2004, 02:25 AM, said:

dako, on Nov 5 2004, 09:57 AM, said:

now, I am trying beta gal. Promega 1:200, 1:1000, 1:2000 I use just 1 reagent from M.O.M kit - Blocking reagent, preincubation with Normal Goat Serum and secondary antibody I take Goat anti Mouse Peroxidase (Sigma), so I am curiouse, When I will have result I will write you. Where are zou come from? And which language do you speak? Bye

For using the Promega antibody, I also used M.O.M. kit, including blocking reagent and working solution. After that I chose ABC kit for secondary antibody of anti-mouse peroxidase. But the difference is very small. Please tell me your result. Good luck!
I am from Japan, I'm a second grade graduate school student at the department of Genetics. Good bye.

Hallo, so result is really bad, negative control and all dilutions had backround, it was nearly same, so without positivities. But I have good expeariens with Chemicon and Sanes, problem was that same activity was on double transgenic and also wild type mouse! Realy nice positivities! Negative controls were negative. I am from Czech Republic, I am student of medicine - health technician.. If you have problem with ELSE antibody, write. Have nice day

[kostianchik]

Hi! I'm looking for beta-gal protocols for paraffin sections staining. (human tissue). Could u tell me- is it possible? Where should I look at it? Thanks guys.