Recombinant Anti-CD133 antibody [EPR20980-104] (ab216323)
![Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-451724-anti-cd133-antibody-epr20980-104-western-blot-caco2-hela.jpg "Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20980-104] to CD133
- Suitable for: WB, IHC-P, Flow Cyt, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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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
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Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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.
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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: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20980-104 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry reagents
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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 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.
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| IHC-P | (2) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Flow Cyt |
1/100.
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| IP |
1/70.
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| Notes |
|---|
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WB
1/1000. Predicted molecular weight: 97 kDa. |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Flow Cyt
1/100. |
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IP
1/70. |
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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Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323)All lanes : Anti-CD133 antibody [EPR20980-104] (ab216323) at 1/1000 dilution
Lane 1 : Caco-2 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 20 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 minutesab216323 was shown to react with CD133 in western blot. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab216323 overnight at 4°C at a 1 in 1000 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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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-287525-anti-cd133-antibody-epr20980-104-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (ab216323)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). -
![Flow Cytometry - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-287521-anti-cd133-antibody-epr20980-104-flow-cytometry.jpg)
Flow Cytometry - Anti-CD133 antibody [EPR20980-104] (ab216323)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.
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![Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-287528-anti-cd133-antibody-epr20980-104-western-blot.jpg)
Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323)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 minutesBlocking: 5% NFDM/TBST.
Negative control: HEK-293T (PMID 20167130).
The MW observed is consistent with the literature (PMID 20717901; PMID 18645205).
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![Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-287527-anti-cd133-antibody-epr20980-104-western-blot.jpg)
Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323)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
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-287524-anti-cd133-antibody-epr20980-104-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (ab216323)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). -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-287523-anti-cd133-antibody-epr20980-104-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (ab216323)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). -
![Immunoprecipitation - Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-287519-anti-cd133-antibody-epr20980-104-immunoprecipitation.jpg)
Immunoprecipitation - Anti-CD133 antibody [EPR20980-104] (ab216323)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. -
![Anti-CD133 antibody [EPR20980-104] (ab216323)](https://www.abcam.com/ps/products/216/ab216323/Images/ab216323-8-benefits-of-recombinant-antibodies.png)
Anti-CD133 antibody [EPR20980-104] (ab216323)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (30)
ab216323 has been referenced in 30 publications.
- Shang Y et al. MiR-7-5p/KLF4 signaling inhibits stemness and radioresistance in colorectal cancer. Cell Death Discov 9:42 (2023). WB ; Human . PubMed: 36732504
- Wang J et al. Functional assessment of miR-1291 in colon cancer cells. Int J Oncol 60:N/A (2022). WB ; Human . PubMed: 34981812
- Cai JH et al. HucMSCs-exosomes containing miR-21 promoted estrogen production in ovarian granulosa cells via LATS1-mediated phosphorylation of LOXL2 and YAP. Gen Comp Endocrinol 321-322:114015 (2022). Flow Cyt ; Human . PubMed: 35271888
- Tang B et al. xCT contributes to colorectal cancer tumorigenesis through upregulation of the MELK oncogene and activation of the AKT/mTOR cascade. Cell Death Dis 13:373 (2022). Flow Cyt ; Human . PubMed: 35440604
- Li L et al. Knockdown of miR-92a suppresses the stemness of colorectal cancer cells via mediating SOCS3. Bioengineered 13:5613-5624 (2022). WB ; Human . PubMed: 35184640