about 'mini-protein 2' by bio-rad ???

D. KIM dkim at NMSU.Edu
Tue Aug 17 17:17:57 EST 1999

I am guessing at what you are doing.  You run multiple lanes of protein
extract, blot and cut out the lanes to incubate with different hybridoma
supernatants.  You can then use a second-antibody system to detect whether
the hybridoma is producing a monoclonal antibody that binds to an
appropriately-sized protein band.  Is this correct?

If so, then you desire to probe a single protein extract with a variety of
different hybridoma supernatants.

There is an apparatus sometimes called a "miniblotter manifold",
consisting of a piece of plastic with machined grooves cut into it.  You
run a gel with a single broad lane containing your target protein extract.
Blot this protein onto a membrane and block.  Next, the membrane is
sandwiched into the manifold.  The grooves create multiple chambers, each
of which can be filled with your hybridoma supernatants (20 - 40 per gel).
After the primary incubation, drain the chambers and wash them.
Disassemble the manifold and proceed with washes and detection.  

I know this is not a clear description, but I hope it will lead you to the
apparatus I have in mind.

Daniel Kim
dkim at nmsu.edu

Joo Yun Hong <jhong1 at uic.edu> wrote:
: hello!!

: i'm using hydridoma supernatant to screen monoclonal antibody about
: protein on a
: membrane. since the amount of supernatant i can use is limited to a
: small volume, i
: used this apparatus named  'mini-protean2'  from the bio-rad. but it has
: been giving
: me a problem of leakage between  the wells. so i was wondering if anyone
: could  tell
: me about similar kind of apparatus using a membrane to screen multiple
: samples of
: hydridoma supernatant  or have  any idea of eliminating the leakage
: problem.

:                                                                    thank
: you

: joo         jhong1 at uic.edu

More information about the Methods mailing list