Anti-MAP4K5 Antibody

• Catalog Number: orb669236
• Category: Antibodies
• Description: Anti-MAP4K5 Antibody. Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human.
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human MAP4K5, which shares 94.4% amino acid (aa) sequence identity with both mouse and rat MAP4K5.
• UniProt ID: Q9Y4K4
• MW: 95 kDa
• Tested applications: ICC, IF, IHC, WB
• Application notes: Western blot, 0.25-0.5μg/ml, Human Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human Immunocytochemistry/Immunofluorescence, 5μg/ml, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Antibody Type: Primary Antibody
• 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.
• Alternative names: Protein NDRG3; N-myc downstream-regulated gene 3 p
• Note: For research use only

IF analysis of MAP4K5 using anti-MAP4K5 antibody. MAP4K5 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-MAP4K5 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.

IHC analysis of MAP4K5 using anti-MAP4K5 antibody. MAP4K5 was detected in paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-MAP4K5 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of MAP4K5 using anti-MAP4K5 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 Jurkat whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human HEK293 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-MAP4K5 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 MAP4K5 at approximately 95 KD. The expected band size for MAP4K5 is at 95 KD.