Anti-CARD4/NOD1 Antibody

• Catalog Number: orb234340
• Category: Antibodies
• Description: Anti-CARD4/NOD1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ICC, IHC, WB
• Reactivity: Human, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human CARD4 recombinant protein (Position: M1-M160). Human CARD4 shares 82% amino acid (aa) sequence identity with mouse CARD4.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 107 kDa
• UniProt ID: Q9Y239
• 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: Nucleotide-binding oligomerization domain-containi
• Note: For research use only
• Application notes: Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/mlImmunocytochemistry, 0.5-1μg/mlWestern blot, 0.1-0.5μg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IHC analysis of CARD4 using anti-CARD4 antibody. CARD4 was detected in immunocytochemical section of Hela 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 1 µg/ml rabbit anti-CARD4 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of CARD4 using anti-CARD4 antibody. CARD4 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 1 µg/ml rabbit anti-CARD4 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of CARD4 using anti-CARD4 antibody. CARD4 was detected in immunocytochemical section of SMMC-7721 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 1 µg/ml rabbit anti-CARD4 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of CARD4 using anti-CARD4 antibody. CARD4 was detected in paraffin-embedded section of Human Placenta Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CARD4 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 CARD4 using anti-CARD4 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: A549 Whole Cell Lysate, Lane 2: Rat Cardiac Muscle Tissue Lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-CARD4 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 1:10000 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 CARD4 at approximately 107 KD. The expected band size for CARD4 is at 107 KD.