2013 Promega catalogue

Cell Signaling
Product Size Cat.# Price ($)
Anti-ACTIVE ® CaM KII pAb, Rabbit, (pT 286 ) 40 μl V1111 678
For Research Use Only. Not for Use in Diagnostic Procedures.
kDa
250 –
98 –
64 –
50 –
36 –
30 –
16 –
6 –
4 –
0.5µg 2.5µg 5µg 12.5µg 25µg
+ – + – + – + – + –
1 2 3 4 5 6 7 8 9 10 11
– Phospho
T 286
CaM KII
Detection of CaM KII by Anti-ACTIVE ® CaM KII pAb in Western
analysis of rat brain homogenate. Lanes 2, 4, 6, 8 and 10 contain
autophosphorylated (+) brain cytosolic protein in the amounts shown; lanes
3, 5, 7, 9 and 11 contain nonphosphorylated (–) brain cytosolic protein in
the amounts shown. The presence of the autophosphorylated CaM KII was
detected using the Anti-ACTIVE ® CaM KII pAb diluted 1:5,000.
Panel A
Panel B
Panel C
2055TA02_8A
Immunocytochemical detection of active JNK enzyme in Drosophila
pupal retina using the Anti-ACTIVE ® JNK pAb. Drosophila pupal retina
at 25% of pupal development were fixed in 3% paraformaldehyde in PBS.
The Anti-ACTIVE ® JNK pAb was diluted 1:100 in PBS containing 10% fetal
bovine serum and 0.2% Triton ® X-100. Samples were incubated with
the primary antibody overnight at 4˚C, washed 3 times (10 minutes each)
with 0.2% Triton ® X-100 and then incubated with a goat anti-rabbit Cy ® 3
conjugate for 2 hours at 4˚C. Whole mounts were visualized with a Zeiss ®
Axioskop fluorescent microscope. The results illustrate the presence of
dually phosphorylated, active forms of JNK in discrete structures of the
fly retinal ommatidia including intense staining of the inner cone cells
as well as the mechanosensory bristles and surrounding pigment cells.
The pattern of staining (which was distinct from results obtained with an
antibody for active p38) and the absence of staining in control experiments
(data not shown) support the high specificity of the Anti-ACTIVE ® JNK pAb.
Image kindly provided by David T. Miller and Ross Cagan, Department of
Molecular Biology and Pharmacology, Washington University School of
Medicine, St. Louis, Missouri.
1825TA07_7A
4
Cell Signaling
Immunocytochemical detection of autophosphorylated CaM KII in
PC12 cells with Anti-ACTIVE ® CaM KII pAb. PC12 cells were adhered
to slides coated with collagen, fixed in 10% paraformaldehyde for 30
minutes, rinsed in PBS and permeabilized in methanol for 10 minutes
at –20˚C. The cells were then blocked in 1% BSA in PBS for 45 minutes
followed by 2% horse serum in PBS for 60 minutes. Cells were incubated
overnight at 4˚C with (Panel A) Ab alone, (Panel B) Ab preincubated
with a nonphosphorylated CaM KII peptide fragment (1µg/ml) or (Panel
C) Ab preincubated with a phosphorylated CaM KII peptide fragment
(1µg/ml). The Anti-ACTIVE ® CaM KII pAb was used at 1:500 dilution and
preincubated with peptide for 8 hours at 4˚C. After incubation with the
Anti-ACTIVE ® CaM KII pAb or Ab/peptide mixture, the cells were rinsed in
PBS and incubated with a donkey anti-rabbit FITC-conjugated secondary
Ab (1:500) for 60 minutes at room temperature. Staining was visualized
with a Zeiss ® fluorescence microscope. The results demonstrate that
preincubation of the Anti-ACTIVE ® CaM KII pAb with phosphorylated
CaM KII peptide completely abolishes immunostaining (Panel C), but
preincubation with nonphosphorylated CaM KII peptide has no effect on
immunostaining (Panel A versus Panel B). Protocols developed and
performed at Promega.
2057TA02_8A
Product Size Cat.# Price ($)
Anti-ACTIVE ® MAPK pAb, Rabbit, (pTEpY) 40 μl V8031 678
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Size Cat.# Price ($)
Anti-ACTIVE ® p38 pAb, Rabbit, (pTGpY) 100 μl V1211 678
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Size Cat.# Price ($)
Anti-ACTIVE ® JNK pAb, Rabbit, (pTPpY) 40 μl V7931 678
120 μl V7932 1127
For Research Use Only. Not for Use in Diagnostic Procedures.
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