| TiterZyme® Kits
(ELISA Kits) |
IMPORTANT: We have
recently released a new Total ERK1/2 TiterZyme® EIA kit. Due to
the supply of a critical raw material, this assay kit has been
re-developed using a new detection antibody. Effective
immediately, the new format Total ERK EIA kit (900-152) will be
replacing the older version (900-103).
The specifications for the kit have changed slightly and are
outlined in the attached document:
total ERK Technical Bulletin
 |
|
 |
| ERK 1/2
Colorimetric (EIA) Kits |
| Assay Format |
Catalog # |
Kit Insert |
MSDS |
Assay Layout |
| phospho: 96 Well Kit |
900-098 |
|
|
|
| total: 96 Well Kit |
900-152 |
|
|
|
| ERK
Chemiluminescent (CLIA) Kits |
| Assay Format |
Catalog # |
Kit Insert |
MSDS |
Assay Layout |
| phospho: 96 Well Kit |
910-098 |
|
|
|
|
|
FOR RESEARCH PURPOSES ONLY, NOT FOR DIAGNOSTIC USE. |
 
|
| |
"Quantitative
Determination of Phosphorylated Mitogen-Activated Protein Kinase (pERK)
by Colorimetric and Chemiluminescent Immunometric Assays" (697 KB) -
poster presented at AACR 2003 in Washington, D.C.
"Supersensitive
Quantitative Measurement of Phosphorylated Mitogen-Activated Protein
Kinase by Chemiluminescent Immunoassay" (1 MB) - poster presented at
SBS 2003 in Portland, OR
Stable, Color-Coded Liquid Reagents
|
The mitogen-activated protein kinase (MAPK) pathway exists in all
eukaryotes. It consists of several subgroups including ERK 1/2 (extracellular
signal-regulated kinase),
JNK and
p38 kinases. These kinases regulate and control fundamental cellular
processes including proliferation, apoptosis, survival and
differentiation. They are activated by diverse stimuli including
cytokines, growth factors, irradiation, changes in osmolarity, heat and
sheer stress.
MAP kinases are characterized by their requirement for dual
phosphorylation for full activation. ERK1 (44 kDa) and ERK2 (42 kDa)
kinases are characterized by their requirement for dual phosphorylation
at a conserved T-E-Y motif . The literature contains numerous and
extensive reviews on MAP kinase.
|
|
 |
1. Pipet Standards and Samples in duplicate into the wells and incubate.
2. Aspirate and wash the plate.
3. Pipet yellow Antibody into the wells and incubate.
4. Aspirate and wash the plate.
5. Pipet blue Conjugate into the wells and incubate.
6. Aspirate and wash the plate.
7. Pipet Substrate into the wells and incubate.
8. EIA: Pipet Stop Solution and read on a plate reader at 450 nm. CLIA:
Read in a suitable luminometer or luminescence detector.
9. Calculate sample concentrations from Standard Curve.
|
| |
phospho ERK EIA
(900-098) |
total ERK EIA
(900-152) |
phospho ERK CLIA
(910-098) |
| RANGE |
62.5-2,000 pg/mL
|
156-20,000 pg/mL
|
37.5-600 pg/mL
|
| SAMPLE SIZE |
100 µL |
100 µL |
100 µL |
| SAMPLES PER 96
WELL KIT |
40 in duplicate
|
38 in duplicate
|
41 in duplicate
|
| SENSITIVITY |
38.3 pg/mL
|
22.9 pg/mL
|
34.4 pg/mL
|
| PRECISION |
4.3-6.4% Intra
4.9-11.5% Inter |
1.7-4.5% Intra
5.8-10.4% Inter |
5.2-11.3% Intra
11.4-14.2% Inter |
SAMPLE TYPE
recommended dilution |
RIPA cell lysis buffer
( 1:4)
|
cell lysates
|
RIPA cell lysis buffer
(1:4)
|
| TIME TO ANSWER |
3 Hours |
3 Hours |
3 Hours |
| SPECIES
SPECIFICITY |
non-specific
|
non-specific
|
non-specific
|
|
WARNING: If the end user chooses to not use the provided RIPA Cell Lysis
Buffer, it is up to the end user to determine the appropriate dilution
of samples and assay validation for their chosen cell lysis buffer.
|
| Compound |
Cross Reactivity |
Compound |
Cross Reactivity |
| phospho ERK
|
| phospho ERK |
100% |
|
|
| <0.01%:
non-phosphorylated ERK / phospho p38 / non-phosphorylated p38 /
phospho JNK / non-phosphorylated JNK / phospho AKT /
non-phosphorylated AKT |
| Compound |
Cross Reactivity |
Compound |
Cross Reactivity |
| total ERK
EIA |
| ERK1, active |
100% |
ERK2, active |
100% |
| ERK1, inactive |
100% |
ERK2, inactive |
92.6% |
| <0.08%: MEK1, inactive
/ MEK2, inactive / p38 alpha, inactive / JNK alpha1alpha,
inactive |
| Compound |
Cross Reactivity |
Compound |
Cross Reactivity |
| phospho ERK
CLIA |
| bioactive phospho ERK |
100% |
|
|
| <0.01%:
non-phosphorylated ERK / phospho p38 / non-phosphorylated p38 /
phospho-JNK / non-phosphorylated JNK / phospho-AKT /
non-phosphorylated AKT |
|
+32 (0) 16
58 90 45 
+32 (0) 16
50 90 45