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StripObuffer (Antibody Stripping Buffer)
(Cat #:
FGI
1989w and
FGI
-1989s)
Product Specification Sheet
Product
Description
The
idea of reusing protein blots derives strength from the fact that
antigen‑antibody complexes (e.g., immunoaffinity columns)
can be disrupted and the solid phase immunoaffinity column reused
several times. However,
commonly employed elution conditions in immunoaffinity techniques
(low or high pH, use of chaotropic agents) have not been very
effective in stripping antibodies from protein blots.
Fabgennix Int. Inc., has now developed a specially
formulated water based buffer solution that can effectively and
almost quantitatively strip antibodies from its antigens in 5-10
min. at room temperature.
Introduction
Western
blotting is a commonly used technique to study protein structure
and function. Typically, protein samples are electrophoresed on
SDSPAGE‑gels and transferred to a solid support
(nitrocellulose or PVDF membranes) for subsequent probing with
specific antibodies. Unlike advances made in RNA/
DNA
blotting, it has been difficult to strip antibodies from the
complex of its antigen, and reuse blots. Innumerable usage of same
protein blots for probing with different antibodies. That will not
only save investigator’s precious antigen/protein samples that
are not available in abundance, but save invaluable time and
resources also.
Some
of the advantages of re-probing the western blot membranes are:
(i) quantitative
western blotting and quantifying the inter-lane comparision of
samples on the same gel by probing the sdame blot with a house
keeping protein like actin, synaptopohysin etc.
(ii) Comparison
of phosphorylated and nonphosphorylated state of the same protein
on the same blot which will eliminate the sample application and
loading variability (iii) Stripping
buffer also offers the opportunity to investigators to reconfirm
their protein expression results by reprobing the same stripped
blot utilizing a different set of antibodies raised against the
same or different epitope for the same protein or procured by
other vendors.
Our
StripObuffer guarantees multiple stripping on the same blot with
little or no loss to the proteins on the membranes, the
antigenicity is maintained throughout several cycles of stripping.
No heating and No pungent smell of powerful reducing agents
(β-mercaptoethanol andor Dithiotheritol).
Blots could be very effectively stripped off its antibodies
at least 10 times, and it is proven to work on both Nitrocellulose
and PVDF membranes.
Dilute (
1:10
)
the supplied buffer in distilled water prior to use.
Rinse the membrane in
distilled water first and then in stripping buffer at least once
(multiple rinses with buffer results in better signal to noise
ratios and appearance of sharp high contrast bands on
autoradiograms)
Immerse the membrane
completely (at least in 10 ml for each mini-gel membrane) in 1X
stripping buffer.
Gently shake the membrane on
a rocker for 5 min. at room temperature.
Rinse the membrane with
distilled water and then with TBST or with the buffer in which
your primary antibody is diluted.
Proceed for blocking and rest
of the western blotting processes.
Storage:
The reagent can be stored at 4oC for up to 6 months with
out any significant loss in the potency of the reagent.
Related
items:
BindObuffer
(Antibody/protein binding buffer)
Cat. # FGI-1920/1921
200 ml/500 ml
Protease
inhibitor cocktail (Cat # FGI-1943ab)
100 ul
EluObuffer,
Antibody Elution buffer (Cat #
FGI
-1931)
200ml
Secondary
Antibody AP Conjugate (goat anti-mouse IgG, A, M)
250 ul
Alkaline
Phosphatase substrate buffer (5X)
Cat. # FGI-1964
100 ml
\\* For users who
may require more than 5 or more of StripObuffer, please inquire
about bulk material discounts from FabGennix International Inc..
This Product is for Research Use Only and is NOT intended for use in
humans or clinical diagnosis.
062606-0010SF1001Z-rev10.00
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