Overview

  • Product name

    Anti-CD133 antibody [EPR20980-104]
    See all CD133 primary antibodies
  • Description

    Rabbit monoclonal [EPR20980-104] to CD133
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Non human primates
  • Immunogen

    Recombinant fragment within Human CD133 aa 500-800. The exact sequence is proprietary.
    Database link: O43490

  • Positive control

    • WB: Caco-2 and NCCIT whole cell lysate. Human fetal kidney lysate. IHC-P: Human mammary gland, kidney and pancreas tissue. Flow Cytometry: HEK-293T cells. IP: Caco-2 whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab216323 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Predicted molecular weight: 97 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/100.
IP 1/70.

Target

  • Function

    May play a role in cell differentiation, proliferation and apoptosis (PubMed:24556617). Binds cholesterol in cholesterol-containing plasma membrane microdomains and may play a role in the organization of the apical plasma membrane in epithelial cells. During early retinal development acts as a key regulator of disk morphogenesis. Involved in regulation of MAPK and Akt signaling pathways. In neuroblastoma cells suppresses cell differentiation such as neurite outgrowth in a RET-dependent manner (PubMed:20818439).
  • Tissue specificity

    Isoform 1 is selectively expressed on CD34 hematopoietic stem and progenitor cells in adult and fetal bone marrow, fetal liver, cord blood and adult peripheral blood. Isoform 1 is not detected on other blood cells. Isoform 1 is also expressed in a number of non-lymphoid tissues including retina, pancreas, placenta, kidney, liver, lung, brain and heart. Found in saliva within small membrane particles. Isoform 2 is predominantly expressed in fetal liver, skeletal muscle, kidney, and heart as well as adult pancreas, kidney, liver, lung, and placenta. Isoform 2 is highly expressed in fetal liver, low in bone marrow, and barely detectable in peripheral blood. Isoform 2 is expressed on hematopoietic stem cells and in epidermal basal cells (at protein level). Expressed in adult retina by rod and cone photoreceptor cells (at protein level).
  • Involvement in disease

    Retinitis pigmentosa 41
    Cone-rod dystrophy 12
    Stargardt disease 4
    Retinal macular dystrophy 2
  • Sequence similarities

    Belongs to the prominin family.
  • Post-translational
    modifications

    Isoform 1 and isoform 2 are glycosylated.
    Acetylation at Lys-225, Lys-257 and Lys-264 by NAT8 and NAT8B may control PROM1 protein expression and its function in cell apoptosis.
  • Cellular localization

    Apical cell membrane. Cell projection, microvillus membrane. Cell projection, cilium, photoreceptor outer segment. Endoplasmic reticulum. Endoplasmic reticulum-Golgi intermediate compartment. Found in extracellular membrane particles in various body fluids such as cerebrospinal fluid, saliva, seminal fluid and urine.
  • Information by UniProt
  • Database links

  • Alternative names

    • AC133 antibody
    • Antigen AC133 antibody
    • CD133 antibody
    • CORD12 antibody
    • Hematopoietic stem cell antigen antibody
    • hProminin antibody
    • MCDR2 antibody
    • MSTP061 antibody
    • OTTHUMP00000217744 antibody
    • OTTHUMP00000217745 antibody
    • OTTHUMP00000217746 antibody
    • PROM1 antibody
    • PROM1_HUMAN antibody
    • Prominin I antibody
    • Prominin like 1 antibody
    • Prominin like protein 1 precursor antibody
    • Prominin mouse like 1 antibody
    • Prominin-1 antibody
    • Prominin-like protein 1 antibody
    • Prominin1 antibody
    • PROML1 antibody
    • RP41 antibody
    • STGD4 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling CD133 with ab216323 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP), ready to use. Apical/endoluminal staining on ducts of human pancreas, and negative on Langerhans cells of the islets (PMID: 18261235) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP), ready to use.
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-CD133 antibody [EPR20980-104] (ab216323) at 1/1000 dilution

    Lane 1 : Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 97 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking: 5% NFDM/TBST.

    Negative control: HEK-293T (PMID 20167130).

    The MW observed is consistent with the literature (PMID 20717901; PMID 18645205).

  • All lanes : Anti-CD133 antibody [EPR20980-104] (ab216323) at 1/1000 dilution

    Lane 1 : NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate
    Lane 2 : Human fetal kidney

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
    Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 97 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?



    Blocking: 5% NFDM/TBST.

    Exposure time:

    Lane 1: 30 seconds

    Lane 2: 1 minute

  • Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling CD133 with ab216323 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP), ready to use. Apical/endoluminal staining on ducts of human mammary gland (PMID: 18261235) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP), ready to use.
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD133 with ab216323 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous staining on parietal layer of Bowman’s capsule of human kidney (PMID: 19092120) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP), ready to use.
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Flow cytometric analysis of HEK-293T (human embryonic kidney epithelial cell)(Left) / Caco-2 (human colorectal adenocarcinoma epithelial cell)(Right) cell line labeling CD133 with ab216323 at 1/100 (Red) compared with Isotype Control Rabbit monoclonal IgG (ab172730) (BLack) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

    Negative control: 293T (PMID 20167130). Total viable cells were gated for the FC image.

  • CD133 was immunoprecipitated from 0.35 mg Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate with ab216323 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab216323 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution
    Lane 1: Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10 μg (Input).
    Lane 2: ab216323 IP in Caco-2 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of  ab216323 in Caco-2 whole cell lysate (-).
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 1 second.

References

This product has been referenced in:

  • Xu XF  et al. BMX-ARHGAP fusion protein maintains the tumorigenicity of gastric cancer stem cells by activating the JAK/STAT3 signaling pathway. Cancer Cell Int 19:133 (2019). WB ; Human . Read more (PubMed: 31130822) »
  • Lu R  et al. Inhibition of CD133 Overcomes Cisplatin Resistance Through Inhibiting PI3K/AKT/mTOR Signaling Pathway and Autophagy in CD133-Positive Gastric Cancer Cells. Technol Cancer Res Treat 18:1533033819864311 (2019). WB, Flow Cyt ; Human . Read more (PubMed: 31405336) »
See all 6 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6.0
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 23 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 30 2017

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