Anti-OLFM4 Antibody

• Catalog Number: orb546347
• Category: Antibodies
• Description: Anti-OLFM4 Antibody. Tested in ELISA, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived human OLFM4 recombinant protein (Position: D91-Q510).
• UniProt ID: Q6UX06
• MW: 75 kDa
• Tested applications: ELISA, IHC, WB
• Application notes: Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml ELISA, 0.1-0.5μg/ml. 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: Olfactomedin-4; OLM4; Antiapoptotic protein GW112;
• Note: For research use only

IHC analysis of OLMF4 using anti-OLMF4 antibody. OLMF4 was detected in paraffin-embedded section of human colon cancer tissues. 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-OLMF4 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.

IHC analysis of OLMF4 using anti-OLMF4 antibody. OLMF4 was detected in paraffin-embedded section of human rectal cancer tissues. 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-OLMF4 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 OLMF4 using anti-OLMF4 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 placenta tissue lysates Lane 2: human U2OS whole cell lysates Lane 3: human U-87MG whole cell lysates Lane 4: human HL-60 whole cell lysates Lane 5: human K562 whole cell lysates Lane 6: human THP-1 whole cell lysates. 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-OLMF4 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 OLMF4 at approximately 75KD. The expected band size for OLMF4 is at 57KD.

Western blot analysis of OLMF4 using anti-OLMF4 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: rat spleen tissue lysates Lane 2: rat kidney tissue lysates Lane 3: mouse kidney tissue lysates Lane 4: mouse lung tissue lysates Lane 5: mouse SP20 whole cell lysates. 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-OLMF4 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 OLMF4 at approximately 75KD. The expected band size for OLMF4 is at 57KD.