PI4K2B Rabbit Polyclonal Antibody

SKU: orb1819393

Description

Anti-PI4K2B Antibody. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human, Rat.

Research Area

Signal Transduction

Images & Validation

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

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: E.coli-derived human PI4K2B recombinant protein (Position: M1-K475). Human PI4K2B shares 78.4% and 79% amino acid (aa) sequence identity with mouse and rat PI4K2B, respectively
• Target: Phosphatidylinositol 4-kinase type 2-beta
• Molecular Weight: 50 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 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

PI 4 Kinase type 2 beta; PI4K2B; PI4KII BETA; PI4KIIB; PIK42B

Flow Cytometry analysis of MCF-7 cells using anti-PI4K2B antibody. Overlay histogram showing MCF-7 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-PI4K2B 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 (Red line) was also used as a control.

IF analysis of PI4K2B using anti-PI4K2B antibody. PI4K2B was detected in an immunocytochemical section of A549 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-PI4K2B Antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The tissue section was developed using Phalloidin-iFluor 488 Conjugated. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of PI4K2B using anti-PI4K2B 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 30 ug of sample under reducing conditions. Lane 1: human HepG2 whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: rat liver 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-PI4K2B 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 PI4K2B at approximately 50 kDa. The expected band size for PI4K2B is at 55, 50 kDa.

Quick Database Links

Gene Symbol

Phosphatidylinositol 4-kinase type 2-beta

UniProt

Q8TCG2