Monoclonal Antibodies for Apoptosis, Inflammation &
Degenerative Neurological Disorder research
Anti Phosphorylated ASK1 / Anti ASK1
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Mitogen-activated protein (MAP) kinase cascades are activated in response to various extracellular stimuli,
including growth factors and environmental stresses. A MAP kinase kinase kinase (MAPKKK), termed ASK1, was
identified that activated two different subgroups of MAP kinase kinases (MAPKK), SEK1 (or MKK4) and
MKK3/MAPKK6 (or MKK6), which in turn activated stress-activated protein kinase (SAPK, also known as JNK;
c-Jun amino-terminal kinase) and p38 subgroup of MAP kinases, respectively. Overexpression of ASK1 induced
apoptotic cell death, and ASK1 was activated in cells treated with tumor necrosis factor-alpha (TNF-alpha).
Moreover, TNF-alpha-induced apoptosis was inhibited by a catalytically inactive form of ASK1. ASK1 may be a
key element in the mechanism of stress- and cytokine-induced apoptosis.
■ Anti Phosphorylated ASK1, Monoclonal Antibody (Wako Cat. No. 017-22351 (50 µg))
Features:
Reacts with human ASK1 including phosphorylated mouse Thr838 and human Thr845.
Cross-reacts to ASK2 and ASK3, which phosphorylated site are the same as that of ASK1.
No cross-reaction with human ASK1 Thr838 Ala mutant.
Clone No.: PA214
Working Dilution
Western blot
1 : 1,000
Specificity of Anti Phosphorylated ASK1 (PA214)
ASK1 tagged with Flag was transfected into HEK293A cells and Western blot analysis was performed using
anti-phosphorylated ASK1 antibody (PA214). As a result, the band could be detected only in the Stim lane containing
ASK1 overexpressing cells stimulated with H2O2 for phosphorylation.
(Data were provided by Dr. Maruyama, J. and
Noguchi, T., Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo)
(-) lane: Control vector
TA lane: Human ASK1, mutated to Ala at Thr838, which
is the recognition site for phosphorylation
antibody, was overexpressed.
WT lane: Wild type human ASK1 was overexpressed.
Stim lane:   Wild type human ASK1 was overexpressed and
H2O2 stimulation was conducted for activation.
■ Anti ASK1, Monoclonal Antibody (Wako Cat. No. 010-22341 (50 µg))
Features:
Reacts with human and mouse ASK1
No cross-reactive with ASK2.
Clone No.: TC003
Specificity of Anti ASK1 (TC003)
Bone marrow-derived macrophages were extracted from ASK1 (+/+) and ASK1 (-/-) mice and Western blot analysis
was performed using ASK1 antibody (TC003). As a result, samples derived only from ASK1 (+/+) mice could be
detected. (Data were provided by Dr. Maruyama, J. and Noguchi, T., Cell Signaling, Graduate School of
Pharmaceutical Sciences, the University of Tokyo)
References:
1.

2.
Ichijo, H., Nishida, E., Irie, K., et al.:
SCIENCE, 275, 90-94 (1997)
Tobiume, K., Saitoh, M., Ichijo, H., et al.: 
J. Cell Physiol., 191, 95-104 (2002)
Wako Catalog No.
Description
Grade
Package Size
017-22351
Anti Phosphorylated ASK1, Monoclonal
Antibody (Clone No. PA214)
for
Immunochemistry
50 µg
010-22341
Anti ASK1, Monoclonal Antibody
(Clone No. TC003)
50 µg
Desired Antibody Diluent to enhance immunoreaction at Western blotting, Dot blotting and ELISA
Immuno-enhancer
Immuno-enhancer, which is an antibody diluent to enhance
immunoreaction at Western blotting, dot blotting and ELISA
aids in obtaining high signal-to-noise(S/N) ratio.
This kit consists of ready-to-use solutions such as
Reagent A as the diluent of the primary antibody, and
Reagent B as the diluent of the secondary antibody diluent.
Features
■ Enhances Immunoassay Signals, effectively
■ High S/N Ratio
■ Applicable to Ready-to-Use antibody diluent
A549 cell Lysate 5 µg (1) or 10 µg (2) were
subjected to SDS-PAGE and blotted onto
nitrocellulose membranes.
They were blocked and Western blot.
3% skim milk in TBS-T solution was used as the
control.
Primary Antibody: Anti EB1, Rabbit (1:500), for 2
hr-reaction.
Secondary Antibody: HRP conjugated Anti rabbit IgG
(1:7,000), for 1 hr-reaction; Exposure time: 10 sec.
Description
Wako Catalog No.
Package Size
Antibody Diluent to enhance immunoreaction
Immuno-enhancer
<for Blotting>
294-68601
290-68603
298-68604
for 2 tests
for 10 tests
for 40 tests
Listed products are intended for laboratory research use only, and not to be used for drug, food or human use.
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