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Anti-ACTIVE® MAPK pAb, Rabbit, (Cat.# V8031)
is an affinity-purified
polyclonal antibody that specifically recognizes the dually phosphorylated, active form of
MAPK (also known as p44/ERK1 and p42/ERK2) enzymes. Anti-ACTIVE® MAPK pAb is
raised against a dually phosphorylated peptide sequence representing the catalytic core of
the active ERK enzyme. The phosphorylated amino acid residues correspond to Thr183 and
Tyr185 of the p42/ERK2 enzyme. The recommended dilution of Anti-ACTIVE® MAPK
pAb for Western blot analysis is 1:5,000.
- Specificity: Preferentially detects the dually phosphorylated, active
form of the mitogen-activated protein kinase (MAPK) enzymes (ERK1 and ERK2).
- Immunogen: Dually phosphorylated Thr/Glu/Tyr region (pTEpY) derived
from the catalytic core of the active form of the mitogen-activated protein kinase (MAPK)
enzymes, ERK1 and ERK2, corresponding to the Thr183 and Tyr185 of the mammalian ERK2
enzyme.
- Antibody Form: Affinity-purified rabbit IgG; supplied
in PBS (pH 7.4).
- Value: When used at the recommended 1:5,000 dilution, this product will
generate 200ml of blotting solution, sufficient for 20 Western blots of 10ml each.
The Catalog Number of the current Anti-ACTIVE® MAPK is V8031. Prior
to this, the Anti-ACTIVE® MAPK pAb catalog number was V6671. A
brief history of the change is below.
Promega Product Information Sheet (.pdf, 32kb): 
Western Blotting
Protocol:
- Technical Bulletin #TB262: Anti-ACTIVE®
MAPK, p38 and JNK Polyclonal Antibodies and Anti-ACTIVE® Qualified Secondary
Antibody Conjugates.—Contains Promega's optimized protocol for Western analysis of
extracts and has different optimized protocols for PVDF membranes or nitrocellulose
membranes.
Technical Literature:
- Jarvis, B.W. and O'Brien, M. (1999) PC12 Extracts for use with new Anti-pT183
MAPK and Anti-ACTIVE® Antibodies. Promega
Notes 72, 10–12.—Western blots of new NGF-treated and
untreated PC12 Cell Extracts (Cat.# V8110).
- Jarvis, B.W. and Moravec, R. (1998) New Anti-ACTIVE® MAPK & 'pan ERK 1/2'
Antibodies for Western Analysis. Promega
Notes 69, 9–10.—Western blots of NGF-treated PC12 cell
extracts.
- Using Anti-ACTIVE® MAPK, p38 and JNK pAbs for Western blotting. Neural Notes 18, 12–13
Citations
for Western Blot Analysis
Immunocytochemistry
Protocol:
- Technical Bulletin #TB262: Anti-ACTIVE®
MAPK, p38 and JNK Polyclonal Antibodies and Anti-ACTIVE® Qualified Secondary
Antibody Conjugates–Contains Promega's optimized protocol for immunocytochemical
analysis of MAPK in PC12 cells treated with nerve growth factor.
Technical Literature:
- Ahn, N.G., et al. (1999) U0126: An Inhibitor of MKK/ERK Signal Transduction in
Mammalian Cells. Promega Notes 71,
4–7.—ICC of NIH3T3 cells for ERK activation +/– U0126 MEK Inhibitor (Cat.#
V1121).
- Immunostaining with the new Anti-ACTIVE® p38 Antibody. Neural Notes 17, 2–4.—ICC
of NGF-treated PC12 cells.
Citations
for Immunocytochemistry
Immunohistochemistry
Technical Literature:
- Hoffman, R., O'Brien, M. and Jarvis, B.W. (2001) Immunohistochemical staining using
Promega Anti-ACTIVE® and Apoptosis Antibodies. Cell Notes 1, 4–7.—;IHC
of rat brain hippocampus with the Anti-ACTIVE® MAPK pAb and Anti-ERK 1/2 pAb.
Full protocols are provided.
Citations
for Immunohistochemistry
Cross-Reactivity Citations
For information regarding species cross-reactivity, please select one of the
following species for a list of citations:
The Change From V6671 to V8031
In 1996 we introduced the original Anti-ACTIVE® MAPK
(Cat.# V6671). V6671 was a highly concentrated
antibody requiring a 1:20,000 dilution for use in Western blotting applications,
and thus customer received a very small quantity of antibody for making the
Western blotting solutions. Western blotting is the most commonly used method to
analyze MAPK signaling pathways. Many customers contacted Promega and asked that
the antibody be supplied at a lower concentration so dilutions would be easier
and more accurate. In 1998, we decided to make this change in response to our
customer's needs and changed the concentration of the antibody to where only a
1:5,000 dilution was necessary to prepare the blotting solutions. We thought the
change was large enough to require a new catalog number and thus V8031 was born.
You can read more about this change in Promega
Notes 69. Nothing has changed with the immunogen from V6671 to V8031. The
V8031 actually has a more strict Quality Control procedure than the original
V6671. The V6671 only had to pass a Quality Control check for Western blotting.
The V8031 must pass the Western blotting Quality Control check and an
Immunocytochemistry Quality Check as well. To our knowledge this is the most
strict quality control procedure for any anti-dually phosphorylated Erk antibody
available.
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