Anti-CD133/Prom1 Antibody Picoband®

Western blot analysis of CD133/Prom1 using anti-CD133/Prom1 antibody (A01767-4).
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: mouse kidney tissue lysates,
Lane 2: rat kidney 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 rabbit anti-CD133/Prom1 antigen affinity purified polyclonal antibody (Catalog # A01767-4) at 0.25 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 CD133/Prom1 at approximately 130 kDa. The expected band size for CD133/Prom1 is at 130 kDa.

Analysis of tumor infiltrating lymphocytes by Flowcytometer showed that a population of Th1 cells (CD3 + CD4 + CD25 − ) was significantly increased inside tumors of 1-MT, 1-MT+TL and 1-MT+DC-TL treated groups in comparison to PBS ( A , B ). Meanwhile, Treg + cells (CD25 + FOXP3 + ) were significantly reduced in comparison to PBS. Cancer stem cells (CSC) CD133 + viability in tumor microenvironments of all tested groups showed that 1-MT, 1-MT+TL and 1-MT+DC-TL extremely reduced CD133 viability in comparison to PBS group ( C , D ), as well as the analysis of relative mRNA showed that CD133 gene expression was significantly declined in 1-MT+TL and 1-MT+DC-TL treated groups in comparison to PBS group ( E ). These results indicated that 1-MT significantly enhanced TL to elicit immunosurveillance that recognizes and effectively reduces CSC prognosis, which inhibits tumor growth and tumorgensis. ***( P < 0.0001), and **( P < 0.001).
Index in PubMed under a CC BY license. PMID: 29959375

Analysis of tumor microenvironments’ mediators, markers and signaling pathways under the effect of 1-MT-TL/DCs-TL regimens by western blot presented that TGF-β, PDL-1, and NF-κβ2 were significantly inhibited in comparison to PBS ( A ). Meanwhile, IDO, β-catenin, and CD133 showed significant reduction under the effect of 1-MT, 1-MT+TL, and 1-MT+DCs-TL in comparison to PBS ( B ). These results clearly evidenced that 1-MT extremely prohibited main tumor growth, progression, and immune escaping pathways, which activates immune surveillance restore and recruits cellular and humoral immune responses triggering tumor tissues.
Index in PubMed under a CC BY license. PMID: 29959375

IHC analysis of CD133/Prom1 using anti-CD133/Prom1 antibody (A01767-4).
CD133/Prom1 was detected in a paraffin-embedded section of mouse kidney 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-CD133/Prom1 Antibody (A01767-4) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

IHC analysis of CD133/Prom1 using anti-CD133/Prom1 antibody (A01767-4).
CD133/Prom1 was detected in a paraffin-embedded section of rat kidney 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-CD133/Prom1 Antibody (A01767-4) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.