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  1. Link

    cd133-antibody-epr16508-ab222782.pdf

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Neuroscience Cell Type Marker Neural Stem Cell marker
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD133 antibody [EPR16508] (ab222782)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question References (15)

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Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)
  • Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
  • Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
  • Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)
  • Anti-CD133 antibody [EPR16508] (ab222782)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16508] to CD133
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Recombinant Human CD133 protein (ab160218)
Primary
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Anti-CD133 antibody [EPR20980-104] - BSA and Azide free (ab226484)

View more associated products

Overview

  • Product name

    Anti-CD133 antibody [EPR16508]
    See all CD133 primary antibodies
  • Description

    Rabbit monoclonal [EPR16508] to CD133
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

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

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1, Caco-2 (Human colorectal adenocarcinoma epithelial cell), NCCIT (Human pluripotent embryonic carcinoma epithelial cell) and HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate. Human fetal pancreas and fetal kidney lysate. IHC-P: Human pancreas and kidney tissue. Human bladder cancer tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR16508
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neural Stem Cell marker
    • Stem Cells
    • Neural Stem Cells
    • Surface Molecules
    • Stem Cells
    • Hematopoietic Progenitors
    • Surface Molecules
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-CD133 antibody [EPR16508] - BSA and Azide free (ab232642)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human CD133 protein (ab160218)
  • Related Products

    • Anti-CD133 antibody [mAbcam27699] (ab27699)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab222782 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
1/2000. Predicted molecular weight: 97 kDa.
IHC-P (1)
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/2000. 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.

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.
  • Target information above from: UniProt accession O43490 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8842 Human
    • Omim: 604365 Human
    • SwissProt: O43490 Human
    • Unigene: 614734 Human
    • 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

    • Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      All lanes : Anti-CD133 antibody [EPR16508] (ab222782) at 1/2000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : PROM1 knockout HAP1 cell lysate

      Lysates/proteins at 40 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 97 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?


      Exposure time: 8 minutes


      ab222782 was shown to react with CD133 in wild-type HAP1 cells in western blot. Loss of signal was observed when PROM1 knockout sample was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab222782 overnight at 4°C at a 1 in 2000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1/5000 for 1 hour at room temperature before development with ECL reagent and imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)

      Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling CD133 with ab222782 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP)(ready use). Apical/endoluminal staining of human pancreatic ducts, and negative on Langerhans cells of the islets (PMID: 18261235). Counter stained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP)(ready to use).

    • Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      All lanes : Anti-CD133 antibody [EPR16508] (ab222782) at 1/10000 dilution

      Lane 1 : Caco-2 (Human colorectal adenocarcinoma epithelial cell), whole cell lysate
      Lane 2 : HT-29 (Human colorectal adenocarcinoma epithelial cell), whole cell lysate
      Lane 3 : HeLa (Human cervix adenocarcinoma 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

      Predicted band size: 97 kDa


      Exposure time: 5 seconds


      Negative control: HeLa (PMID 9389721).

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

      Blocking buffer: 5% NFDM/TBST.

    • Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      Anti-CD133 antibody [EPR16508] (ab222782) at 1/10000 dilution + NCCIT (Human pluripotent embryonic carcinoma epithelial cell), whole cell lysate at 20 µg

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

      Predicted band size: 97 kDa


      Exposure time: 15 seconds


      Blocking buffer: 5% NFDM/TBST.

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

    • Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      Western blot - Anti-CD133 antibody [EPR16508] (ab222782)
      All lanes : Anti-CD133 antibody [EPR16508] (ab222782) at 1/2000 dilution

      Lane 1 : Human fetal pancreas lysate
      Lane 2 : Human fetal kidney lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 97 kDa


      Exposure time: 3 minutes


      Blocking buffer: 5% NFDM/TBST.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)

      Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD133 with ab222782 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ready to use). Membranous staining on parietal layer of Bowman’s capsule and apical staining on renal tubules in human kidney (PMID: 19092120). Counter stained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ready to use).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR16508] (ab222782)

      Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling CD133 with ab222782 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ready to use). Membranous staining on human bladder cancer tumor cells (PMID: 22899341). Counter stained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ready to use).

    • Anti-CD133 antibody [EPR16508] (ab222782)
      Anti-CD133 antibody [EPR16508] (ab222782)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (15)

    Publishing research using ab222782? Please let us know so that we can cite the reference in this datasheet.

    ab222782 has been referenced in 15 publications.

    • Zhou L  et al. Effects of CD133 expression on chemotherapy and drug sensitivity of adenoid cystic carcinoma. Mol Med Rep 25:N/A (2022). WB ; Human . PubMed: 34791507
    • Ding D  et al. Circ_0007385 regulates cell proliferation, apoptosis and stemness via targeting miR-493-3p/RAB22A axis in non-small cell lung cancer. Thorac Cancer N/A:N/A (2022). WB ; Human . PubMed: 34989145
    • Zhong Y & Lan J Overexpression of Eukaryotic translation initiation factor 3D induces stem cell-like properties and metastasis in cervix cancer by activating FAK through inhibiting degradation of GRP78. Bioengineered 13:1952-1961 (2022). WB, IHC-P ; Human . PubMed: 35104170
    • Li H  et al. Connexin32 regulates expansion of liver cancer stem cells via the PI3K/Akt signaling pathway. Oncol Rep 48:N/A (2022). WB ; Human . PubMed: 35894130
    • Chen Z  et al. The role of LINC01419 in regulating the cell stemness in lung adenocarcinoma through recruiting EZH2 and regulating FBP1 expression. Biol Direct 17:23 (2022). WB ; Human . PubMed: 36050791
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD133 antibody [EPR16508]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Carl Hobbs

    Verified customer

    Submitted Nov 30 2017

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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