Anti-DHODH Antibody Picoband® (monoclonal, 2G7)

Western blot analysis of DHODH using anti-DHODH antibody (M04035-1).
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 Hela whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat testis tissue lysates,
Lane 6: rat liver tissue lysates,
Lane 7: mouse testis tissue lysates,
Lane 8: mouse liver tissue 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 mouse anti-DHODH antigen affinity purified monoclonal antibody (Catalog # M04035-1) 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-mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for DHODH at approximately 43 kDa. The expected band size for DHODH is at 43 kDa.

IHC analysis of DHODH using anti-DHODH antibody (M04035-1).
DHODH was detected in a paraffin-embedded section of human breast cancer 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 mouse anti-DHODH Antibody (M04035-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of DHODH using anti-DHODH antibody (M04035-1).
DHODH was detected in a paraffin-embedded section of human gastric cancer 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 mouse anti-DHODH Antibody (M04035-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of DHODH using anti-DHODH antibody (M04035-1).
DHODH was detected in a paraffin-embedded section of human liver cancer 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 mouse anti-DHODH Antibody (M04035-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of DHODH using anti-DHODH antibody (M04035-1).
DHODH was detected in a paraffin-embedded section of human placenta 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 mouse anti-DHODH Antibody (M04035-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of DHODH using anti-DHODH antibody (M04035-1).
DHODH was detected in a paraffin-embedded section of human renal carcinoma 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 mouse anti-DHODH Antibody (M04035-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

M2c macrophages increase ferroptosis resistance in gastric cancer cells. a The CCK-8 method was used to detect the survival of gastric cancer cells (Hgc27 and MKN45) intervened with RSL3 for 24 h. b The CCK-8 method was used to detect the survival of gastric cancer cells (Hgc27 and MKN45) intervened with Fer-1 for 24 h. c The expression of SOD in different intervention groups. d The expression of MDA in different intervention groups. e The expression of GSH in different intervention groups. f The expression of TGFβ1 protein WB in different cell lines. g The expression results of TGFβ1 protein. h The expression of key ferroptosis proteins WB in different cell lines. i The expression results of FSP1 protein. j Expression results of DHODH protein. k Expression results of GPX4 protein. l SLC7A11 protein expression results. m The intervention of RSL3 on the expression of key ferroptosis protein WB in different co culture groups. n The expression results of GPX4 protein. o SLC7A11 protein expression results. p The WB expression of key proteins involved in ferroptosis in different intervention groups. q The expression results of GPX4 protein. r The expression results of SLC7A11 protein. s Fluorescence results of mitochondrial membrane potential in different intervention groups. Scale bar=50 μm. *p<0.05, **p<0.01, ***p<0.001.
Index in PubMed under a CC BY license. PMID: 39991579