Anti-MFSD12 Antibody Picoband®

Western blot analysis of MFSD12 using anti-MFSD12 antibody (A16855-1).
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 SiHa whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human PC-3 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-MFSD12 antigen affinity purified polyclonal antibody (Catalog # A16855-1) 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 1:5000 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 MFSD12 at approximately 52 kDa. The expected band size for MFSD12 is at 52 kDa.

The protein expression analysis of MFSD12. (A) Pan-cancer protein expression profile of MFSD12 and representative IHC staining of tissue microarrays in HPA database. (B) IHC analysis of MFSD12 in LIHC tumor tissues and paired adjacent non-tumor liver tissues. (C) Quantification of immunostains for MFSD12 by IOD analysis. * P < 0.05, ** P < 0.01. IHC, immunohistochemistry; HPA, Human Protein Atlas; LIHC, liver hepatocellular carcinoma; IOD, integrated optical density;.
Index in PubMed under a CC BY license. PMID: 41194934

The knockdown of MFSD12 inhibited the proliferation, migration, and invasion of LIHC cells, as well as the TIM-3/Galectin-9 signaling pathway. (A, B) RT-qPCR and Western blot validation of MFSD12 silencing efficiency using siRNAs (si-MFSD12–1 to −4) with GAPDH as loading control. (C) CCK-8 cell viability assay showing reduced HEP 3B2.1–7 cells proliferation after MFSD12 knockdown (si-MFSD12-3). (D) Transwell assay revealed a reduction in the migratory and invasive capabilities of HEP 3B2.1–7 cells following the knockdown of MFSD12. (E) Immunoblot analysis of EMT markers and TIM-3 axis components showing up-regulation of E-cadherin and down-regulation of Vimentin, MMP-2, MMP-9, HAVCR2 (TIM-3) and LGALS9 in si-MFSD12-treated cells. * P < 0.05, ** P < 0.01, *** P < 0.001. CTRL, control untreated; si-NC, negative control siRNA; si-MFSD12, MFSD12-targeting siRNA; E-cadherin, epithelial cadherin; MMP-2/9, matrix metalloproteinase-2/9; HAVCR2, hepatitis A virus cellular receptor 2 (TIM-3); LGALS9, lectin galactoside-binding soluble 9 (Galectin-9).
Index in PubMed under a CC BY license. PMID: 41194934

IHC analysis of MFSD12 using anti-MFSD12 antibody (A16855-1).
MFSD12 was detected in a paraffin-embedded section of human testicular seminoma 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-MFSD12 Antibody (A16855-1) 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.

IHC analysis of MFSD12 using anti-MFSD12 antibody (A16855-1).
MFSD12 was detected in a paraffin-embedded section of human colon adenocarcinoma 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-MFSD12 Antibody (A16855-1) 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.