Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16508 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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 |
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WB |
1/2000. Predicted molecular weight: 97 kDa.
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IHC-P | (1) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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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
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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
modificationsIsoform 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
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Database links
- Entrez Gene: 8842 Human
- Omim: 604365 Human
- SwissProt: O43490 Human
- Unigene: 614734 Human
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Alternative names
- AC133 antibody
- Antigen AC133 antibody
- CD133 antibody
see all
Images
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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 minutesab222782 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.
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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).
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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 secondsNegative control: HeLa (PMID 9389721).
The MW observed is consistent with the literature (PMID 20717901; PMID 18645205).
Blocking buffer: 5% NFDM/TBST.
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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 secondsBlocking buffer: 5% NFDM/TBST.
The MW observed is consistent with the literature (PMID 20717901; PMID 18645205).
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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 minutesBlocking buffer: 5% NFDM/TBST.
The MW observed is consistent with the literature (PMID 20717901; PMID 18645205).
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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).
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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).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (28)
ab222782 has been referenced in 28 publications.
- Shen Y et al. CDK5RAP2 is a Wnt target gene and promotes stemness and progression of oral squamous cell carcinoma. Cell Death Dis 14:107 (2023). IHC-P ; Human . PubMed: 36774351
- Nong S et al. HN1L promotes stem cell-like properties by regulating TGF-β signaling pathway through targeting FOXP2 in prostate cancer. Cell Biol Int 46:83-95 (2022). PubMed: 34519127
- Hou D et al. Circular RNA circASPM promotes the progression of glioblastoma by acting as a competing endogenous RNA to regulate miR-130b-3p/E2F1 axis. J Cancer 13:1664-1678 (2022). PubMed: 35371308
- Tiong TY et al. Targeting the SREBP-1/Hsa-Mir-497/SCAP/FASN Oncometabolic Axis Inhibits the Cancer Stem-like and Chemoresistant Phenotype of Non-Small Cell Lung Carcinoma Cells. Int J Mol Sci 23:N/A (2022). PubMed: 35806291
- Bi Y et al. CD133, but Not CD44, May Serve as a Novel Biomarker for Differential Diagnosis Between Basal Cell Carcinoma and Trichoblastomas. Clin Cosmet Investig Dermatol 15:1517-1526 (2022). PubMed: 35941854