Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > General Forum > Molecular Biology News and Announcements > Instructions for Posting, Help, and Frequently Asked Questions
Register Search Today's Posts Mark Forums Read

Instructions for Posting, Help, and Frequently Asked Questions Instructions for Posting, Help, and Frequently Asked Questions. If you aren't sure how to start a new thread or post messages check here!


Instructions to Post a Reply.

Instructions to Post a Reply. - Instructions for Posting, Help, and Frequently Asked Questions

Instructions to Post a Reply. - Instructions for Posting, Help, and Frequently Asked Questions. If you aren't sure how to start a new thread or post messages check here!


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 02-03-2006, 04:12 PM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Cool Instructions to Post a Reply.



To post a reply to a post, simply:

1) Click The Post Reply Button
2) Or simply go down a bit after the posts and fill in the Quick Reply Form. Click on the button "post quick reply"

Cheers
Reply With Quote
  #2  
Old 07-29-2006, 05:15 PM
Pipette Filler
Points: 1,817, Level: 26 Points: 1,817, Level: 26 Points: 1,817, Level: 26
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2006
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Instructions to Post a Reply.

Hello all,
I'm a master's student in a Biology Departmant. I'm facing a trouble in pre-processing blood samples before using them in Western Blot technique. My aim is to examin the expression of Cathpsin-K, i.e. to measure (approx.) the serum level of this enzyme.
So if anybody can provide me with tips, I would be so pleased.
Best Regards.
Reply With Quote
  #3  
Old 07-30-2006, 06:19 AM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: Instructions to Post a Reply.

Quote:
Originally Posted by Mounir
Hello all,
I'm a master's student in a Biology Departmant. I'm facing a trouble in pre-processing blood samples before using them in Western Blot technique. My aim is to examin the expression of Cathpsin-K, i.e. to measure (approx.) the serum level of this enzyme.
So if anybody can provide me with tips, I would be so pleased.
Best Regards.
Dear Mounir,
Welcome to the forum! you have posted in a place that few will be able to help, but I can move your post somewhere else and also help you here.

I assume Cathpsin-K is in the serum? not the blood cells in that case...

As you are using Blood, you would need to:

1) centrifuge the samples (you would need a large amount to begin with)

2) isolate the serum by taking only the supernatant (top layer)

3) it would be difficult to blot so you could load a large amount (or precipitate a larger amount of serum into a smaller volume and load that)

4) blot for that protein

is this a good guide or did I miss anything?

Reply With Quote
  #4  
Old 07-31-2006, 10:53 AM
Pipette Filler
Points: 1,817, Level: 26 Points: 1,817, Level: 26 Points: 1,817, Level: 26
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2006
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Response to the instructions

Dear admin.,
First of all I would like to tell you that I'm pleased about your care. The steps you mentioned are themselves the ones I proceeded with. I was centrifuging the blood sample, isolating the serum, and then loading (5) or (10) microliters of the serum after mixing it with loading buffer. The former amount yeilded better bands in the gel but these are still so jammed in which you can't make a reading. I guess if we can get rid of certain kinds of proteins which are known to be found in the serum in high amounts, this may be a solution!!

Please advice,
Regards.
Reply With Quote
  #5  
Old 08-05-2006, 12:10 PM
Pipette Filler
Points: 1,817, Level: 26 Points: 1,817, Level: 26 Points: 1,817, Level: 26
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2006
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Response to the Instructions.

Dear admin.,
First of all, I would like to tell you that I'm pleased about your care. The steps you mentioned are themselves the ones I proceeded with. I was centrifuging the blood samples, isolating the serum, and then loading (5) or (10) microliters of the serum after mixing it with loading buffer. The former amount yielded better bands on the gel but these are still so jammed that you can name them a smear rather than separate bands.

Please advice,
Regards
Reply With Quote
  #6  
Old 08-10-2006, 11:13 PM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: Instructions to Post a Reply.

It seems your antibodies are not very good (as you are getting many many bands - not good), it would help to optimize also the blotting procedure. You should take a look in the forum and this website how to optimize western blots.

Also, post the western blot picture so that we can help you more,

admin
Reply With Quote
The Following User Says Thank You to admin For This Useful Post:
D.raqad (09-04-2011)
  #7  
Old 09-08-2010, 06:11 PM
Pipette Filler
Points: 507, Level: 10 Points: 507, Level: 10 Points: 507, Level: 10
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Instructions to Post a Reply.

Hi,

I've tried posting before. So I'm trying again.

Cheers,
Ken
Reply With Quote
Reply

Tags
instructions , post , reply


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
How to Post and Reply in the Molecular Forum moleculardude Molecular Biology News and Announcements 51 12-27-2010 09:17 AM
POSTopec Fusione Fredda REFLUOPETROLIO LEVItazione MAGNETICA MagLEVscorie mafia CFR KO Global Democrazia TRIVOLUZIONE Era Ora ! CavalLETTA jerecuccata e fuga in SOL DO maggiore Tripoli HAMMANET cameragni CFR mafiaassirobabilonese comuscisti LEVIMOLCAatYAHOOGROUPSdotCOM FrancoLeviATtiscaliDOTit Forum Chimica 0 09-29-2009 12:19 PM
The Theory of Moving Dimensions: The Time Dimension is Moving Relative to The Three Spatial Dimensions Captain Ranger McCoy Physics Forum 14 07-03-2005 03:14 AM
Sci.chem FAQ - Part 1 of 7 Bruce Hamilton Chemistry Forum 0 01-15-2004 09:06 AM
MAKE EASY MONEY FAST !!! Adwithers2 Chemistry Forum 0 08-11-2003 04:43 AM


All times are GMT. The time now is 10:54 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 0.18035 seconds with 16 queries