Anti-Human PROM1 Therapeutic 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.

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ICC

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.

For immunocytochemistry, cells were either cell surface labeled in the cold (A) or permeabilized with saponin after fixation (B). As negative controls, only the secondary antibody was used, and cells were counterstained with DAPI (A, B). Scale bars, 30 μm

Thamm, K., Graupner, S., Werner, C., Huttner, W. B., & Corbeil, D. (2016). Monoclonal antibodies 13A4 and AC133 do not recognize the canine ortholog of mouse and human stem cell antigen prominin-1 (CD133). PloS one, 11(10).

IB

Figure 2 The commercial anti-human and anti-mouse antibodies fail to detect canine prominin-1 by immunoblotting.

(A-C) Detergent lysates prepared from MDCK cells stably transfected with human or mouse prominin-1, canine prominin-1-GFP as well as wild type cells (MDCK) were analyzed by SDS-PAGE under reducing (A, C) and non-reducing (B) conditions and immunoblotting using mAbs AC133, AC141, 293C3, and 13A4 or polyclonal antibody against GFP. β-actin and α-tubulin were used as loading controls. As negative controls, only the secondary antibody (as indicated) was used (C). (D) Lysates from canine prominin-1-GFP transfected cells were incubated with (+) or without (-) PNGase F prior to immunoblotting with anti-GFP antibody or others (as indicated). The arrow and open arrowhead indicate the plasma membrane-associated form and endoplasmic reticulum-associated form of prominin-1, respectively. Asterisk indicates potential disulfide-bridged prominin-1 dimers or multimers, and the black arrowhead shows deglycosylated prominin-1. Molecular mass markers (kDa) are indicated.

Thamm, K., Graupner, S., Werner, C., Huttner, W. B., & Corbeil, D. (2016). Monoclonal antibodies 13A4 and AC133 do not recognize the canine ortholog of mouse and human stem cell antigen prominin-1 (CD133). PloS one, 11(10).

IB

Figure 3 The commercial anti-human and anti-mouse antibodies fail to detect canine prominin-1 by immunoblotting.

Lysates from canine prominin-1-GFP transfected cells were incubated with (+) or without (-) PNGase F prior to immunoblotting with anti-GFP antibody or others. The arrow and open arrowhead indicate the plasma membrane-associated form and endoplasmic reticulum-associated form of prominin-1, respectively. Asterisk indicates potential disulfide-bridged prominin-1 dimers or multimers, and the black arrowhead shows deglycosylated prominin-1. Molecular mass markers (kDa) are indicated.

Thamm, K., Graupner, S., Werner, C., Huttner, W. B., & Corbeil, D. (2016). Monoclonal antibodies 13A4 and AC133 do not recognize the canine ortholog of mouse and human stem cell antigen prominin-1 (CD133). PloS one, 11(10).


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

For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

See other products for "PROM1"

* Abbreviations
3D IHC3D Immunohistochemistry
ActivActivation
AgonistAgonist
ApopApoptosis
BABioassay
BIBioimaging
BlockBlocking
Cell ScreeningCell Screening
SeparationCell Separation
ChIPChromatin Immunoprecipitation
CMCDComplement Mediated Cell Depletion
CostimCostimulation
CytCytotoxicity
DepletionDepletion
DBDot Blot
EMElectron Microscopy
ELISAEnzyme-linked Immunosorbent Assay
ELISPOTEnzyme-linked Immunosorbent Spot
FCFlow Cytometry
FuncSFunctional Assay
GSGel Super Shift Assay
HAHemagglutination
IAImmunoassay
IBImmunoblotting
ICCImmunocytochemistry
IDImmunodiffusion
IFImmunofluorescence
IHCImmunohistochemistry
IHC-FrImmunohistochemistry-Frozen
IHC-PImmunohistochemistry-Paraffin
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
Live cell imagingLive cell imaging
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
MultiplexMultiplex bead-based assay
NeutNeutralization
PPProtein Purification
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
TCTissue Culture
TBTurbidimetry
WBWestern Blot

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