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Product Info Summary
| SKU: | A02354-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-MEKK2/MAP3K2 Antibody Picoband®
SKU/Catalog Number
A02354-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-MEKK2/MAP3K2 Antibody Picoband® catalog # A02354-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, 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-MEKK2/MAP3K2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02354-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human MEKK2/MAP3K2 recombinant protein (Position: M1-Q364).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A02354-1 is reactive to MAP3K2 in Human, Rat
Observed Molecular Weight
78 kDa
Calculated molecular weight
69.7 kDa
Background of MAP3K2
MAP3K2, mitogen-activated protein kinase kinase kinase 2, also known as MEKK2, belongs to the mitogen-activated protein kinase (MAPK) kinase kinase gene family, which is involved in regulating multiple MAPK signaling pathways. Endogenous MAP3K2 localized mainly to the cytosol of unstimulated HeLa cells and rat fibroblasts. The MAP3K2 gene is mapped to chromosome 2 by the International Radiation Hybrid Mapping Consortium. MAP3K2 is a major upstream activator of the JNK, MAPK cascade. MAP3K2 was identifies as 1 of 6 kinases that regulate different steps of the caveolar cycle. Silencing of MAP3K2 strongly reduced kiss-and-run dynamics, leading to an accumulation of caveolar structures at the cell surface without affecting caveolae coat assembly or clustering.
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
A02354-1 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, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, Human
Positive Control
WB: human HEK293 whole cell, human Hela whole cell, human K562 whole cell, human HepG2 whole cell, human MCF-7 whole cell, human Jurkat whole cell, rat kidney tissue
ICC/IF: A431 cell
FCM: SiHa cell
Validation Images & Assay Conditions
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Western blot analysis of MEKK2/MAP3K2 using anti-MEKK2/MAP3K2 antibody (A02354-1).
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 50ug of sample under reducing conditions.
Lane 1: human HEK293 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: human MCF-7 whole cell lysates,
Lane 6: human Jurkat whole cell lysates,
Lane 7: rat kidney tissue 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 rabbit anti-MEKK2/MAP3K2 antigen affinity purified polyclonal antibody (Catalog # A02354-1) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MEKK2/MAP3K2 at approximately 78KD. The expected band size for MEKK2/MAP3K2 is at 78KD.
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IF analysis of MEKK2/MAP3K2 using anti-MEKK2/MAP3K2 antibody (A02354-1).
MEKK2/MAP3K2 was detected in immunocytochemical section of A431 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 rabbit anti-MEKK2/MAP3K2 Antibody (A02354-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
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Flow Cytometry analysis of SiHa cells using anti-MEKK2/MAP3K2 antibody (A02354-1).
Overlay histogram showing SiHa cells stained with A02354-1 (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 rabbit anti-MEKK2/MAP3K2 Antibody (A02354-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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.
Specific Publications For Anti-MEKK2/MAP3K2 Antibody Picoband® (A02354-1)
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