Anti-PDCD5 Antibody

Western blot analysis of PDCD5 using anti-PDCD5 antibody (A02613-1).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: human U251 whole cell lysates,
Lane 4: human HEL whole cell lysates,
Lane 5: human SiHa whole cell lysates,
Lane 6: human Hela whole cell lysates,
Lane 7: human 293T 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-PDCD5 antigen affinity purified polyclonal antibody (A02613-1) at 1:1000 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for PDCD5 at approximately 14 kDa. The expected band size for PDCD5 is at 14 kDa.

IHC analysis of PDCD5 using anti-PDCD5 antibody (A02613-1).
PDCD5 was detected in a paraffin-embedded section of human stomach tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PDCD5 Antibody (A02613-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Methylating effect of daphnetin and 5-aza-dc on DR3, PDCD5, FasL and p53 in CIA rat synovial cells. Four experimental groups consisting of untreated cell control, or treated with 5-aza-dc at 20 μM, daphnetin at 40 μg/mL, or combination of 5-aza-dc (20 μM) and daphnetin (40 μg/mL). Cells were cultured, treated and harvested as described in Materials and Methods. Total DNA was extracted and used for MSP after bisulphite modification. M: methylated amplification products, U: hypomethylated amplification products.
Index in PubMed under a CC BY license. PMID: 25311560

Effect of daphnetin and 5-aza-dc on expression of DR3, PDCD5, FasL, p53 (A) and DNMT1, DNMT3a, DNMT3b (B) in CIA rat synovial cells. Four experimental groups consisting of untreated cell control, or treated with 5-aza-dc at 20 μM, daphnetin at 40 μg/mL, or combination of 5-aza-dc (20 μM) and daphnetin (40 μg/mL). Cells were cultured, treated and harvested as described in Matreials and Methods. Total RNA was extracted and cDNA was synthesized. After reverse transcription, cDNA was used for real time-PCR. Relative quantification of gene expression was performed by the 2-ΔΔCt method. The results show the mean ± S.D. of six independent experiments. ▲P < 0.05 compared with control, ●P < 0.05 compared with 5-aza-dc, ☆P > 0.05 compared with 5-aza-dc, ★P > 0.05 compared with daphnetin, *P < 0.05 compared with daphnetin.
Index in PubMed under a CC BY license. PMID: 25311560

Effect of daphnetin on DR3, PDCD5, FasL and p53 protein expression in CIA rats synovial cells. Cells were cultured, treated and harvested as described Materials and Methods. Four experimental groups consisting of untreated cell control, 5-aza-dc at 20 μM, daphnetin at 40 μg/mL, or combination of 5-aza-dc (20 μM) and daphnetin (40 μg/mL). The results show the mean ± S.D. of six independent experiments. ▲P < 0.05 compared with control, ●P < 0.05 compared with 5-aza-dc, ★P > 0.05 compared with 5-aza-dc, *P < 0.05 compared with daphnetin.
Index in PubMed under a CC BY license. PMID: 25311560