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Product Info Summary
| SKU: | M00996-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-MEK2/MAP2K2 Antibody Picoband® (monoclonal, 2B4)
SKU/Catalog Number
M00996-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-MEK2/MAP2K2 Antibody Picoband® (monoclonal, 2B4) catalog # M00996-2. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-MEK2/MAP2K2 Antibody Picoband® (monoclonal, 2B4) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00996-2)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Monoclonal
Clone Number
2B4
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human MEK2/MAP2K2 recombinant protein (Position: M1-V400).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00996-2 is reactive to MAP2K2 in Human, Mouse, Rat
Observed Molecular Weight
45 kDa
Calculated molecular weight
44.4 kDa
Background of MAP2K2
Dual specificity mitogen-activated protein kinase kinase 2 (MAP2K2), also called PRKMK2 or MEK2, is an enzyme that in humans is encoded by the MAP2K2 gene. The protein encoded by this gene is a dual specificity protein kinase that belongs to the MAP kinase kinase family. MAP2K2 is mapped to 19p13.3. This kinase is known to play a critical role in mitogen growth factor signal transduction, and the inhibition or degradation of this kinase is found to be involved in the pathogenesis of Yersinia and anthrax. Recombinant MEK2 and MEK1 both could activate human ERK1 in vitro, and they further characterized biochemically the 2 MAP2Ks. MAP2K2 has been shown to interact with MAPK3 and ARAF.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M00996-2 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human HEK293 whole cell, human HepG2 whole cell, human Jurkat whole cell, human K562 whole cell, rat brain tissue, rat PC-12 whole cell, mouse brain tissue, mouse Raw2647 whole cell
ICC/IF: MCF-7 cell
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (M00996-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.
Lane 1: human HEK293 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat PC-12 whole cell lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse Raw264.7 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-MEK2/MAP2K2 antigen affinity purified monoclonal antibody (Catalog # M00996-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for MEK2/MAP2K2 at approximately 45KD. The expected band size for MEK2/MAP2K2 is at 45KD.
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Flow Cytometry analysis of A431 cells using anti-MEK2/MAP2K2 antibody (M00996-2).
Overlay histogram showing A431 cells stained with M00996-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- MEK2/MAP2K2 Antibody (M00996-2, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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IF analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (M00996-2).
MEK2/MAP2K2 was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti-MEK2/MAP2K2 Antibody (M00996-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Specific Publications For Anti-MEK2/MAP2K2 Antibody Picoband® (monoclonal, 2B4) (M00996-2)
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