Anti-OIF/Ogn Antibody Picoband®

Western blot analysis of OIF using anti-OIF antibody (A07061-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: recombinant mouse OGN protein 1ng.
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-OIF antigen affinity purified polyclonal antibody (Catalog # A07061-1) 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for OIF at approximately 34KD. The expected band size for OIF is at 34KD.

Culture with OGN-overexpressing CAFs-derived conditioned medium inhibits ovarian cancer cell aggressiveness ( A – B ) The mRNA expression and protein levels of OGN were examined in CAFs and NFs by qRT-PCR and Immunoblotting, respectively. ( C – D ) SKOV3 and A2780 cells were co-cultured with CAFs- or NFs-derived conditioned medium (CAFs-CM/NFs-CM) and examined for the mRNA expression and protein levels of OGN in SKOV3 and A2780 cells by qRT-PCR and Immunoblotting, respectively. ( E – F ) OGN overexpression was achieved in CAFs by transducing OGN overexpression vector (OGN-OE); OGN overexpression was confirmed by qRT-PCR and Immunoblotting, respectively. Then, SKOV3 and A2780 cells were cultured in control medium (con-CM), CAFs-CM, NF-CM, CAFs (lv-NC)-CM, or CAFs (OGN-OE)-CM, and examined for cell viability by MTT assay ( G ); DNA synthesis capacity by EdU ( H ); cell invasion by Transwell with chambers pre-coated with Matrigel ( I ); the protein levels of E-cadherin, vimentin, p-mTOR, mTOR, p-Akt, and Akt by Immunoblotting ( J ). ** P < 0.01, compared with NFs or CAFs (NC) group; ## P < 0.01 compared with CAFs (NC)-CM group
Index in PubMed under a CC BY license. PMID: 38402185

miR-1290 directly targets OGN and inhibits OGN expression ( A ) Targetscan, miRDIP, and literature search methods were used to search for miRNAs that might target OGN and serve as oncogenic miRNAs, and two miRNAs (miR-223-3p and miR-1290) were obtained. ( B ) The expression of miR-223-3p and miR-1290was examined in CAFs and NFs by qRT-PCR. ( C – D ) miR-1290 overexpression or inhibition was achieved in CAFs by transducing miR-1290 mimics or inhibitor; miR-1290 overexpression or inhibition was confirmed by qRT-PCR and Immunoblotting, respectively. ( E ) CAFs were transduced with miR-1290 mimics or inhibitor and examined for the mRNA expression by qRT-PCR. ( F ) Wild- and mutant-type OGN luciferase reporter vectors (wt-OGN/mut-OGN) were constructed as described in the M&M section. Then, wt-OGN or mut-OGN was co-transduced into 293T cells with miR-1290 mimics/inhibitor; the luciferase activity was determined. ( G ) The expression and location of miR-1290 and OGN in CAFs and NFs were determined by FISH assay. miR-1290 (green), OGN (red). Scale bar = 20 μm. * P < 0.05, ** P < 0.01, compared with NC mimics; ## P < 0.01, compared with NC inhibitor
Index in PubMed under a CC BY license. PMID: 38402185

The miR-1290/OGN axis in CAFs modulates ovarian cancer cell aggressiveness SKOV3 and A2780 cells were cultured in CAFs co-transduced with miR-1290 mimics and OGN-OE and examined for cell viability by MTT ( A ); DNA synthesis capacity by EdU ( B ); cell invasion by Transwell with chambers pre-coated with Matrigel ( C ); the protein levels of E-cadherin, vimentin, p-mTOR, mTOR, p-Akt, and Akt by Immunoblotting ( D ). ** P < 0.01, compared with CAFs (NC + NC mimics)-CM group; ## P < 0.01 compared with CAFs (OGN-OE + miR-1290 mimics)-CM group
Index in PubMed under a CC BY license. PMID: 38402185

The miR-1290/OGN axis in CAFs modulates tumorigenic capacity of ovarian cancer cell in vivo Xenograft transplanted tumor models were established in nude mice by injecting a mixture of SKOV3 cells with CAFs or infected CAFs (infected with miR-1290 mimics or lv-OGN) and nude mice were separated into five groups ( n = 6 per group): the SKOV3 cells + CAFs group, the SKOV3 cells + CAFs (lv-NC + NC mimics) group, the SKOV3 cells + CAFs (lv-OGN + NC mimics) group, the SKOV3 cells + CAFs (lv-NC + miR-1290 mimics) group, the SKOV3 cells + CAFs (lv-OGN + miR-1290 mimics) group. ( A ) Images of the tumors in each group. ( B ) Tumor volumes were measured every five days from the 10th day of the experiment. ( C ) Tumor weight were determined at the 30th day. ( D – E ) The mRNA levels of OGN and miR-1290 in mice tumor tissues were detected by qRT-PCR. ( F ) Ki67 and Vimentin levels in tumor tissues were examined using IHC staining. Scale bar = 20 μm. ( G ) The protein levels of E-cadherin, vimentin, p-mTOR, mTOR, p-Akt, and Akt in tumor tissues were determined using immunoblotting. * P < 0.05, ** P < 0.01, compared between SKOV3 cells + CAFs group and SKOV3 cells + CAFs (lv-NC + NC mimics); ## P < 0.01 compared with SKOV3 cells + CAFs (lv-NC + NC mimics) group; && P < 0.01 compared with SKOV3 cells + CAFs (lv-OGN + miR-1290 mimics) group
Index in PubMed under a CC BY license. PMID: 38402185

Sandwich ELISA - Recombinant mouse OIF/Ogn protein standard curve.
Use in combination with reagents from Mouse OIF/Ogn ELISA Kit EZ-Set (DIY Antibody Pairs) (EZ1617).