DAP1/DAP Antibody

SKU: orb763114

Description

Anti-DAP1/DAP Antibody. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.

Images & Validation

• Tested Applications: FC, ICC, IF, IHC, WB
• Reactivity: Human
• Application Notes:
  Western blot, 0.25-0.5 μg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human DAP1/DAP, which shares 92.9% amino acid (aa) sequence identity with both mouse and rat DAP1/DAP.
• Molecular Weight: 15 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
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Platelet basic protein; PBP; C-X-C motif chemokine 7; Leukocyte-derived growth factor; LDGF; Macrophage-derived growth factor; MDGF; Small-inducible cytokine B7; PPBP; CTAP3; CXCL7; SCYB7; TGB1; THBGB1; NAP-2

Flow Cytometry analysis of HL-60 cells using anti-DAP1/DAP antibody. Overlay histogram showing HL-60 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-DAP1/DAP 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 DAP1/DAP using anti-DAP1/DAP antibody. DAP1/DAP was detected in an immunocytochemical section of SiHa 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-DAP1/DAP 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 DAP1/DAP using anti-DAP1/DAP antibody. DAP1/DAP was detected in a paraffin-embedded section of human lung 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-DAP1/DAP 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 DAP1/DAP using anti-DAP1/DAP 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 U-937 whole cell lysates, Lane 2: human RT4 whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: human Hela whole cell lysates, Lane 5: human HepG2 whole cell lysates, Lane 6: human U20S whole cell lysates, Lane 7: human HL-60 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-DAP1/DAP 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 DAP1/DAP at approximately 15 kDa. The expected band size for DAP1/DAP is at 15 kDa.

Quick Database Links

UniProt

P51397