Direct immunodetection within SDS-PAGE GEl? - Is it possible? (Sep/24/2008 )
Dear all,
I would like to know is it possible to conduct the direct immunodetection within SDS-PAGE gel? What are the reasons or factors that make it impossible?
Although I found there are some kits for direct immunodetection within SDS-PAGE gel (Pierce), the gel was pre-treated (Not same as original SDS-PAGE gel). I would like to know the reason behind.
Many thanks.
BC
-peggybee1-
QUOTE (peggybee1 @ Sep 24 2008, 07:20 PM)
Dear all,
I would like to know is it possible to conduct the direct immunodetection within SDS-PAGE gel? What are the reasons or factors that make it impossible?
Although I found there are some kits for direct immunodetection within SDS-PAGE gel (Pierce), the gel was pre-treated (Not same as original SDS-PAGE gel). I would like to know the reason behind.
Many thanks.
BC
I would like to know is it possible to conduct the direct immunodetection within SDS-PAGE gel? What are the reasons or factors that make it impossible?
Although I found there are some kits for direct immunodetection within SDS-PAGE gel (Pierce), the gel was pre-treated (Not same as original SDS-PAGE gel). I would like to know the reason behind.
Many thanks.
BC
I don't know the mechanism behind the kit.
I think we don't normally do direct immunodetection on original SDS-PAGE gel due to protein diffusion in the gel.
Hope this may help.
-Minnie Mouse-
QUOTE
I think we don't normally do direct immunodetection on original SDS-PAGE gel due to protein diffusion in the gel.
And also the protein is three dimensional spread in the gel, if you transfer it to a membran, it is "flat" in two dimensions, so the antibody has easier access to a bigger surface of protein. Antibodies can not invade the gel and bind efficiently to the protein in the PAGE-matrix.
I don't know this kit, you have to tell us more so that we can conclude how it works.
-biomaus-
QUOTE (biomaus @ Sep 24 2008, 11:30 PM)
QUOTE
I think we don't normally do direct immunodetection on original SDS-PAGE gel due to protein diffusion in the gel.
And also the protein is three dimensional spread in the gel, if you transfer it to a membran, it is "flat" in two dimensions, so the antibody has easier access to a bigger surface of protein. Antibodies can not invade the gel and bind efficiently to the protein in the PAGE-matrix.
I don't know this kit, you have to tell us more so that we can conclude how it works.
I don't agree with you. When the proteins were treated with SDS and merca,.all proteins should be denatured and became primary in structure...
-peggybee1-
QUOTE (peggybee1 @ Sep 24 2008, 06:20 PM)
Dear all,
I would like to know is it possible to conduct the direct immunodetection within SDS-PAGE gel? What are the reasons or factors that make it impossible?
Although I found there are some kits for direct immunodetection within SDS-PAGE gel (Pierce), the gel was pre-treated (Not same as original SDS-PAGE gel). I would like to know the reason behind.
Many thanks.
BC
I would like to know is it possible to conduct the direct immunodetection within SDS-PAGE gel? What are the reasons or factors that make it impossible?
Although I found there are some kits for direct immunodetection within SDS-PAGE gel (Pierce), the gel was pre-treated (Not same as original SDS-PAGE gel). I would like to know the reason behind.
Many thanks.
BC
you need much longer incubation times as the gel matrix hinders the diffusion of antibodies into the gel...
-The Bearer-
QUOTE
I don't agree with you. When the proteins were treated with SDS and merca,.all proteins should be denatured and became primary in structure...
I was not referring to the protein 2- or 3-dimensional structure but the distribution all over the gel matrix. And as the gel has a depth of 0.75 cm or 1cm, the proteins are not distributed in 2 dimensions but 3. Sure they should be in primary structure.
-biomaus-