Anti-Transferrin/TF Antibody Picoband®

Comparison of the binding ability of PVDF membrane and NC membrane to medium molecular weight proteins. ( a ) The pooled sera proteins (0.1–3.0 μg) were subjected to 8% SDS-PAGE. The electroblotted membranes are PVDF membrane (up) and NC membrane (down), respectively. The membranes were incubated with anti-alpha-1-acid glycoprotein (AGP), anti-eukaryotic transformation extension factor 1 alpha 2 (EEF1A2) and anti-transferrin (TF) antibodies. ( b ) Staining intensities were statistically analyzed (n = 3 individual experiments). Pink bar, PVDF membrane; Blue bar, NC membrane. Band intensities were analyzed and compared using Image Lab software (Bio-Rad Laboratories) and GraphPad Prism version 6. N.S., not significant. All values are means ± S.E. (error bars).
Index in PubMed under a CC BY license. PMID: 34103620

Western blot analysis of Transferrin using anti-Transferrin antibody (PB9827).
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 placenta tissue lysates,
Lane 2: human hepatocellular carcinoma tumor tissue (HCCT) lysates,
Lane 3: human hepatocellular carcinoma paracancerous tissue (HCCP) lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse Hepa1/6 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-Transferrin antigen affinity purified polyclonal antibody (Catalog # PB9827) 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 Transferrin at approximately 77 kDa. The expected band size for Transferrin is at 77 kDa.

IHC analysis of Transferrin using anti-Transferrin antibody (PB9827).
Transferrin was detected in a paraffin-embedded section of human esophageal 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 rabbit anti-Transferrin Antibody (PB9827) 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.

Flow Cytometry analysis of HepG2 cells using anti-Transferrin antibody (PB9827).
Overlay histogram showing HepG2 cells stained with PB9827 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Transferrin Antibody (PB9827, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.