MEKK2/MAP3K2 Antibody

SKU: orb669234

Description

Anti-MEKK2/MAP3K2 Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Rat.

Research Area

Cardiovascular Research, Cell Biology, Neuroscience

Images & Validation

• Tested Applications: FC, ICC, IF, WB
• Dilution range: Western blot, 0.25-0.5μg/ml, Human, Rat Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x10^6 cells, Human ELISA, 0.1-0.5μg/ml, Human
• Reactivity: Human, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human MEKK2/MAP3K2 recombinant protein (Position: M1-Q364).
• Target: Mitogen-activated protein kinase kinase kinase 2
• Molecular Weight: 78 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

MAP3K2; MAPK/ERK kinase kinase 2; MAPKKK2; MEK kinase 2; MEKK 2; MEKK2; MEKK2B

Flow Cytometry analysis of SiHa cells using anti-MEKK2/MAP3K2 antibody. Overlay histogram showing SiHa cells (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 (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of MEKK2/MAP3K2 using anti-MEKK2/MAP3K2 antibody. MEKK2/MAP3K2 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-MEKK2/MAP3K2 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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.

Western blot analysis of MEKK2/MAP3K2 using anti-MEKK2/MAP3K2 antibody. 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 50 ug 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 150 mA 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 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 with Tanon 5200 system. A specific band was detected for MEKK2/MAP3K2 at approximately 78 KD. The expected band size for MEKK2/MAP3K2 is at 78 KD.

Quick Database Links

Gene Symbol

Mitogen-activated protein kinase kinase kinase 2

UniProt

Q9Y2U5