Anti-Human PROM1 Recombinant Antibody (AC133)

CAT#: TAB-1074CL

This antibody can bind to Prominin-1 which has ADCC activity and CDC activity. It also can potentially work as a pharmaceutical composition fot treatment of tumors.

Published Data Gene Expression
Figure 1 The MDCK cells stably transfected with human or mouse prominin-1, canine prominin-1-GFP as well as wild type cells (MDCK) were analyzed either by immunocytochemistry using mAbs AC133, AC141, 293C3 and 13A4. Figure 2 The commercial anti-human and anti-mouse antibodies fail to detect canine prominin-1 by immunoblotting. Figure 3 The commercial anti-human and anti-mouse antibodies fail to detect canine prominin-1 by immunoblotting.
Figure 1 Colon Figure 2 Liver Figure 3 Pancreas Figure 4 Kidney Figure 5 RNA cell line category: Cell line enhanced (AF22, CACO-2, CAPAN-2, EFO-21, NTERA-2, RPTEC TERT1)

Specifications

  • Host Species
  • Mouse
  • Specificity
  • Human
  • Clone
  • AC133
  • Applications
  • ELISA, ICC, IB
  • Related Disease
  • Cancers expressing the Prominin-1 protein

Applications

  • 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.

Target

  • 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
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Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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