NCAPH Rabbit Polyclonal Antibody

SKU: orb1743899

Description

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

Research Area

Epigenetics & Chromatin

Images & Validation

• Tested Applications: ELISA, FC, ICC, IF, WB
• Dilution Range: Western blot, 0.1-0.25 μg/ml, Human 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

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human NCAPH recombinant protein (Position: R91-K721).
• Target: Condensin complex subunit 2
• Molecular Weight: 100 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

Mediator of RNA polymerase II transcription subunit 20; Mediator complex subunit 20; TRF-proximal protein homolog; hTRFP; MED20; TRFP

Flow Cytometry analysis of 293T cells using anti-NCAPH antibody. Overlay histogram showing 293T 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-NCAPH 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 NCAPH using anti-NCAPH antibody and anti-Beta Tubulin antibody. NCAPH was detected in immunocytochemical section of A549 cell. 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-NCAPH Antibody and mouse anti-Beta Tubulin antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG and DyLight®488 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of NCAPH using anti-NCAPH 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 Hela whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human K562 whole cell 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-NCAPH antigen affinity purified polyclonal antibody at 0.25 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 NCAPH at approximately 100 kDa. The expected band size for NCAPH is at 83-100 kDa.

Quick Database Links

Gene Symbol

Condensin complex subunit 2

UniProt

Q15003