Anti-Human PROM1 Recombinant Antibody (AC133) (CAT#: TAB-1074CL)

• Host Species
• Mouse

• Specificity
• Human

• Clone
• AC133

• Applications
• ELISA, ICC, IB

• Related Disease
• Cancers expressing the Prominin-1 protein

• Application Notes
• Immunocytochemistry
  Cell surface labeling-Wild type and transfected MDCK cells were grown on fibronectin-coated glass coverslips or in eight-well chambers (μ-Slide 8 Well) until day 6 post-confluence.Cells were rinsed with Ca/Mg buffer (phosphate-buffered saline containing 1 mM CaCl₂ and 0.5 mM MgCl₂) and incubated in blocking buffer (Ca/Mg buffer containing 0.2% gelatin) for 30 min at 4°C. Cells were incubated with either mouse mAb AC133 (epitope CD133/1; 1:50), AC141 (epitope CD133/2; 1:50), 293C3 (CD133/2, 1:50) or rat mAb 13A4 (1:100) diluted in blocking buffer for 1 h at 4°C. Cells were washed with PBS and then fixed in 4% paraformaldehyde (PFA) in PBS for 30 min at room temperature. Coverslips were then rinsed in PBS containing 50 mM NH₄Cl and incubated in this same solution for 10 min.
  Intracellular labeling-PFA-fixed cells were permeabilized and blocked with PBS containing 0.2% gelatin and 0.2% saponin for 30 min at room temperature prior to the incubation with anti-prominin-1 antibodiesfor 30 min at room temperature. To visualize the primary cilium cells were labeled with a mouse mAb anti-acetylated α-tubulin (1:1000, clone 6-11B-1) followed by Alexa546-conjugated goat anti-mouse IgG2b (1:600)for 30 min in permeabilization buffer. For labeling with homemade antibodies, PFA-fixed cells were incubated in SDS buffer (0.005% SDS and 0.2% gelatin in PBS) for 30 min at room temperature and washed with PBS containing 0.2% gelatin for 10 min. Cells were labeled with mouse mAb 80B258 (10 μg/ml) or rabbit antiserum αhE2 (1:500) diluted in PBS containing 0.2% gelatin for 30 min at room temperature. In all cases, Alexa555/546-conjugated goat/donkey anti-mouse, anti-rabbit or anti-rat IgG antibodies (1:500) were used to detect anti-prominin-1 antibodies. Nuclei were counterstained with 4'-6-diamidino-2-phenylindole (DAPI, 1 μg/ml). Samples were washed three times in PBS containing 0.2% gelatin, once in PBS, once in H₂O and mounted in Mowiol 4.88. Cells were observedwith a Zeiss LSM 700 confocal laser scanning microscope.
  Cell detergent extracts were prepared from wild type or transfected MDCK cells (corresponding to one-tenth of a unit of 6-well plate) and separated by SDS-PAGE (7.5%) under reducing or non-reducing conditions and transferred to polyvinylidene difluoride (PVDF) membranes (pore size: 0.45 μm) using a semi-dry transfer cell system. After transfer, membranes were incubated in blocking buffer (PBS containing 0.3% Tween 20 and 5% milk powder) overnight at 4°C. Incubation with primary antibodies against prominin-1; mouse mAb AC133 (1:100), AC141 (1:100), 293C3 (1:100), 80B258 (1:1500) or rat mAb 13A4 (1:500), and rabbit antiserum αhE2 (1:1500) was performed for 1 h at room temperature. The GFP fusion protein was detected using a rabbit anti-GFP polyclonal antibody (1:2000). As a loading control, mouse mAb AC-74 against β-actin (1:10,000) or DM1A against α-tubulin (1:10,000) were used. All antibodies were diluted in blocking buffer. Antigen-antibody complexes were detected using appropriate horseradish peroxidase-conjugated secondary antibodies (1:5000) and visualizedwith enhanced chemiluminescence reagents (ECL system). Membranes were exposed to films.

• Alternative Names
• PROM1; prominin 1; RP41; AC133; CD133; MCDR2; STGD4; CORD12; PROML1; MSTP061; prominin-1; hProminin; antigen AC133; prominin-like protein 1; hematopoietic stem cell antigen

• Gene ID
• 8842

• UniProt ID
• O43490

• ADC-014LCT
• Anti-CD133 (AC133)-vcMMAF ADC

• TAB-1074CL-S(P)
• Anti-Human PROM1 Recombinant Antibody scFv Fragment (AC133)

• TAB-1074CL-F(E)
• Anti-Human PROM1 Recombinant Antibody Fab Fragment (AC133)

• VS-1024-FY265
• Anti-PROM1 (clone AC133) Recombinant Antibody Coupled Liposome (VS-1024-FY265)

Select a product category from the dropdown menu below to view related products.