Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Histology Forum > Immunofluorescence Forum
Register Search Today's Posts Mark Forums Read

Immunofluorescence Forum Immunofluorescence Forum. Talk, discuss and post questions on immunofluorescence techniques including paraffin, frozen sections or cultured cells.


Incubation Time for Immunofluorescence Antibodies?

Incubation Time for Immunofluorescence Antibodies? - Immunofluorescence Forum

Incubation Time for Immunofluorescence Antibodies? - Immunofluorescence Forum. Talk, discuss and post questions on immunofluorescence techniques including paraffin, frozen sections or cultured cells.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 05-21-2008, 04:13 AM
Pipette Filler
Points: 2,212, Level: 30 Points: 2,212, Level: 30 Points: 2,212, Level: 30
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2006
Posts: 10
Thanks: 0
Thanked 0 Times in 0 Posts
Question Incubation Time for Immunofluorescence Antibodies?



Hello what is the optimal incubation time for primary and secondary antibodies for IF (Immunofluorescence)
Reply With Quote
  #2  
Old 05-21-2008, 04:15 AM
Pipette Filler
Points: 2,449, Level: 31 Points: 2,449, Level: 31 Points: 2,449, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jan 2006
Posts: 19
Thanks: 0
Thanked 2 Times in 2 Posts
Default Re: Incubation Time for Immunofluorescence Antibodies?

Hi i will suggest what i am doing myself.

for Primary Antibodies incubation time is about 45 min to 60 minutes.

for Secondary antibodies the incubation time of 30 min to 45 minutes are enough.

Even you can standardize your incubation time within this limit.

However, not that the incubation time will depend on your antibody and antibody concentration, sample, buffers etc.

Depending on your protocol you may need to play around with the conditions a bit.

Reply With Quote
  #3  
Old 07-22-2008, 04:07 AM
Pipette Filler
Points: 366, Level: 7 Points: 366, Level: 7 Points: 366, Level: 7
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2008
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Incubation Time for Immunofluorescence Antibodies?

Hi, I have my protein expressed in pIRS2 vector (EGFP). Now i am interested to see nuclear staining with other molecule. Therefore, I treated with primary antibody and secondry i used Alexa-594 antibody (red). I can see the red nuclear staining. But in green field I saw weared image. Could some one suggest me? I used Flourescence microscope, should I use confocal microscope? Or Can I used the similarly treat with primary antibody and then strained with Alexa 494 (green). In that case, I will have both colours with similar strength. But, I am afraid that EGFP will creat problem. Please give me some suggestion.

Thanks inadvance

Last edited by wasmn; 07-22-2008 at 04:13 AM.
Reply With Quote
  #4  
Old 02-12-2009, 01:36 PM
Pipette Filler
Points: 2, Level: 1 Points: 2, Level: 1 Points: 2, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Incubation Time for Immunofluorescence Antibodies?

hallo Leute ich habe ein problem mit IF! Ich habe ein CA9 antikörper der bestens ergebnisse gibt wenn ich Alexa 488 als sec antikörper verwende. Allerdings die selbe CA9 mit Alexa 350 gibt kein spezifische signal. Die einzige unterschied ist also nur die farbe von alexa sonst nichts. Was könnte ich tun?? Vielen dank! Constantin
Reply With Quote
  #5  
Old 04-02-2009, 09:22 PM
Pipette Filler
Points: 900, Level: 16 Points: 900, Level: 16 Points: 900, Level: 16
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2009
Posts: 7
Thanks: 0
Thanked 1 Time in 1 Post
Default Re: Incubation Time for Immunofluorescence Antibodies?

Quote:
Originally Posted by jiji324 View Post
Hello what is the optimal incubation time for primary and secondary antibodies for IF (Immunofluorescence)
Hi,

we incubate primary antibody over night at 4 degree (conc. 1:300), and secondary 1 hour at room temperature, same concentration..
I have read many protocols that are using the same incubation time.
good luck!
Reply With Quote
The Following User Says Thank You to kraksi For This Useful Post:
admin (04-03-2009)
Reply

Tags
antibodies , immunofluorescence , incubation , time


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Moving Dimensions Theory Book Due Out in Fall 05--Very Rough Draft: 4th Dimensions Expanding Relative to 3 Spatial Dimensions jollyrogership@yahoo.com Physics Forum 64 03-31-2012 10:24 AM
Simply put, MOVING DIMENSIONS THEORY is THE NEW MODEL: http://physicsmathforums.com drelliot@gmail.com Physics Forum 0 08-29-2006 06:32 AM
Moving Dimensions Theory!! Rock On!! drelliot@gmail.com Physics Forum 1 07-06-2006 05:19 PM
Does any of this make sense ??? Yrael Physics Forum 3 09-12-2005 03:11 AM


All times are GMT. The time now is 09:35 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.17244 seconds with 16 queries