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OneMinute®
Western Blot Stripping Buffer
In
ONE minute, your membrane is ready for re-probing
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OneMinute® Western Blot Stripping Buffer,
a revolutionary
stripping buffer, is designed to specifically
strip off antibodies but not other non-antibody proteins. Therefore, not only
the blot can be re-blotting for multiple times,
but also re-blocking is generally not necessary.
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OneMinute®
Western Blot Stripping Buffer immediately
makes your membrane ready for re-blotting in
ONE minute. It
spends only 30 seconds for stripping and 3x 10
seconds for washing.
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Highlights:
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- Faster, easier and more efficient: only
need 1 min (Fig2,
3)
- No
significant loss of transferred
proteins (Fig1, 5)
- Multiple times of re-probes: ~10
times (Fig6)
- Re-blocking is usually not necessary
- Perform at RT, no offensive odor
- Work
for both nitrocellulose and PVDF
membranes (Fig4)
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Protocols:
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1.
Immerse membrane in OneMinute®
Western Blot Stripping Buffer
for 30 sec.
2. Wash the blot with Washing Buffer (TBST or PBST)
3 x 10 seconds*.
*The
blot is ready for re-probing. Re-blocking
membrane is usually not necessary. |
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OneMinute: 30sec at RT, washing 3x 10 sec, No
re-blocking.
Competitor: 15min at 50℃, washing 3x 10min,
re-blocking 1 hour, washing 3x 15min |
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| Fig
1. OneMinute®
prevents the loss of transferred proteins comparing to traditional stripping buffer.
Please click the figure for details. |
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Harsh
stripping buffer (2%SDS, 100mM 2-ME, 50mM Tris,
pH6.8, perform at 50C for 15min) always leads to
significant loss of transferred proteins, which
results in dramatically signal loss in subsequent
re-blotting. When using mild stripping buffer such
as Pierce's Restore, researchers always suffer
incomplete removal of antibodies, especially primary
antibodies. OneMinute®
Western Blot Stripping Buffer represents the
only buffer available to efficiently remove
antibodies (Fig 2, 3, 4), whereas keeping
transferred proteins on the membrane in maximum (Fig
1, 5, 6).
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Fig
2 shows that stripping with
OneMinute®
for
half minute completely
removed both secondary antibodies (panel B) and primary
antibodies to protein 1 (panel C). In contrast,
stripping with Pierce's Restore for 15 minutes failed to strip off
primary antibodies (panel C). This figure demonstrates the superb stripping efficiency of
OneMinute®.
The
ability to strip off high-affinity antibodies by
OneMinute®
is much better than Pierce's Restore (Fig 3). |
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| Fig
2. OneMinute®
is much more efficient than Pierce's Restore.
Please
click the figure for details. |
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3. OneMinute®
is much more efficient than Pierce's Restore.
Please
click the figure for details. |
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4. OneMinute®
woks not only on
nitrocellulose membrane but also on PVDF membrane.
Please
click the figure for details. |
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OneMinute: 10sec at RT, washing 3x 10 sec, No
re-blocking.
Competitor: 15min at 50℃, washing 3x 10min,
re-blocking 1 hour, washing 3x 15min |
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5. Minimal
loss of transferred proteins was achieved by OneMinute®.
Please
click the figure for details. |
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| Fig
6. Multiple
times stripping (10x) by OneMinute®
retained enough transferred proteins on membrane (3rd lane
of panel B).
Please
click the figure for details. |
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| Fig
7. Detect phosphorylated and non-phosphorylated forms of the same
protein by stripping with OneMinute®.
Please
click the figure for details. |
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Related
Products and Links
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Ordering
Information |
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