Western Blot (WB)
(Figure 1. Western blot analysis of PROM1/CD133 using anti-PROM1/CD133 antibody (MBS1753997).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human CACO-2 whole cell lysatesLane 2: human SW620 whole cell lysatesLane 3: mouse kidney tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- PROM1/CD133 antigen affinity purified monoclonal antibody (Catalog # MBS1753997) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PROM1/CD133 at approximately 120KD. The expected band size for PROM1/CD133 is at 120KD. )
Mouse anti-Human, Mouse PROM1/CD133 Monoclonal Antibody | anti-PROM1 antibody
Anti-PROM1/CD133 Antibody (monoclonal, 8B6)
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of PROM1/CD133 using anti-PROM1/CD133 antibody (MBS1753997).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human CACO-2 whole cell lysatesLane 2: human SW620 whole cell lysatesLane 3: mouse kidney tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- PROM1/CD133 antigen affinity purified monoclonal antibody (Catalog # MBS1753997) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PROM1/CD133 at approximately 120KD. The expected band size for PROM1/CD133 is at 120KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of PROM1/CD133 using anti-PROM1/CD133 antibody (MBS1753997).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human CACO-2 whole cell lysatesLane 2: human SW620 whole cell lysatesLane 3: mouse kidney tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- PROM1/CD133 antigen affinity purified monoclonal antibody (Catalog # MBS1753997) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PROM1/CD133 at approximately 120KD. The expected band size for PROM1/CD133 is at 120KD. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of CACO-2 cells using anti-PROM1/CD133 antibody (MBS1753997).Overlay histogram showing CACO-2 cells stained with MBS1753997 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- PROM1/CD133 Antibody (MBS1753997, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of CACO-2 cells using anti-PROM1/CD133 antibody (MBS1753997).Overlay histogram showing CACO-2 cells stained with MBS1753997 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- PROM1/CD133 Antibody (MBS1753997, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Corbeil D, Fargeas CA, Huttner WB (2001). "Rat prominin, like its mouse and human orthologues, is a pentaspan membrane glycoprotein". Biochem Biophys Res Commun 285 (4): 939-44.
3. Shmelkov, S. V., Butler, J. M., Hooper, A. T., Hormigo, A., Kushner, J., Milde, T., St. Clair, R., Baljevic, M., White, I., Jin, D. K., Chadburn, A., Murphy, A. J., Valenzuela, D. M., Gale, N. W., Thurston, G., Yancopoulos, G. D., D'Angelica, M., Kemeny, N., Lyden, D., Rafii, S. CD133 expression is not restricted to stem cells, and both CD133+ and CD133- metastatic colon cancer cells initiate tumors. J. Clin. Invest. 118: 2111-2120, 2008.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a pentaspan transmembrane glycoprotein. The protein localizes to membrane protrusions and is often expressed on adult stem cells, where it is thought to function in maintaining stem cell properties by suppressing differentiation. Mutations in this gene have been shown to result in retinitis pigmentosa and Stargardt disease. Expression of this gene is also associated with several types of cancer. This gene is expressed from at least five alternative promoters that are expressed in a tissue-dependent manner. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Mar 2009]
Uniprot Description
CD133: Binds cholesterol in cholesterol-containing plasma membrane microdomains. Proposed to play a role in apical plasma membrane organization of 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. Interacts with CDHR1 and with actin filaments. 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. Expressed in adult retina by rod and cone photoreceptor cells. Belongs to the prominin family. 7 isoforms of the human protein are produced by alternative splicing.
Protein type: Cell surface; Membrane protein, multi-pass; Membrane protein, integral
Chromosomal Location of Human Ortholog: 4p15.32
Cellular Component: stereocilium; extracellular space; cell surface; photoreceptor outer segment; microvillus membrane; endoplasmic reticulum; integral to plasma membrane; apical plasma membrane; plasma membrane; ER-Golgi intermediate compartment; vesicle; brush border
Molecular Function: actinin binding; protein binding; cadherin binding
Biological Process: photoreceptor cell maintenance; retina morphogenesis in camera-type eye
Disease: Macular Dystrophy, Retinal, 2; Stargardt Disease 4; Retinitis Pigmentosa 41; Cone-rod Dystrophy 12
Research Articles on PROM1
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Product Notes
The PROM1 prom1 (Catalog #AAA1753997) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-PROM1/CD133 Antibody (monoclonal, 8B6) reacts with Human, Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's PROM1/CD133 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Flow Cytometry (FC/FACS/FCM). WB: 0.25-0.5ug/ml|Human, Mouse| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the PROM1 prom1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PROM1/CD133, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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