DAB Peroxidase staining - (Aug/19/2009 )
Hi,
I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?
Thx.
-baboon-
baboon on Aug 19 2009, 11:14 AM said:
Hi,
I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?
Thx.
I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?
Thx.
Check the pH of your pBS it has to be 7.3, I would rather dissolve in pBS only
-pesji-
Thanks,
I shall try dissolving in PBS.....
-baboon-
baboon on Aug 20 2009, 06:14 AM said:
Hi,
I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?
Thx.
I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?
Thx.
Hi there,
I am using the same stuff, but i am using anti mouse HRP, for the DAB I am following the protocol provided by sigma, which is- Crush DAB- (atleast for 1 hour, to make it as a fine powder), try dissolving it in 3-5 ml warm ethanol, finally disolve it in PBS. I would like to discuss more about this, i am from Melbourne and you are?
MS
-miraclestrain-