Western Blot (WB)
(Figure 1. Western blot analysis of Wnt5b using anti-Wnt5b antibody (MBS1753716).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 HEK293 whole cell lysatesLane 2: human SW620 whole cell 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 rabbit anti-Wnt5b antigen affinity purified polyclonal antibody (Catalog # MBS1753716) 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-rabbit 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 # MBS176460) with Tanon 5200 system. A specific band was detected for Wnt5b at approximately 45KD. The expected band size for Wnt5b is at 40KD. )
Rabbit anti-Human Wnt5b Polyclonal Antibody | anti-WNT5B antibody
Anti-Wnt5b Antibody
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of Wnt5b using anti-Wnt5b antibody (MBS1753716).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 HEK293 whole cell lysatesLane 2: human SW620 whole cell 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 rabbit anti-Wnt5b antigen affinity purified polyclonal antibody (Catalog # MBS1753716) 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-rabbit 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 # MBS176460) with Tanon 5200 system. A specific band was detected for Wnt5b at approximately 45KD. The expected band size for Wnt5b is at 40KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of Wnt5b using anti-Wnt5b antibody (MBS1753716).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 HEK293 whole cell lysatesLane 2: human SW620 whole cell 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 rabbit anti-Wnt5b antigen affinity purified polyclonal antibody (Catalog # MBS1753716) 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-rabbit 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 # MBS176460) with Tanon 5200 system. A specific band was detected for Wnt5b at approximately 45KD. The expected band size for Wnt5b is at 40KD. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of U87 cells using anti-Wnt5b antibody (MBS1753716).Overlay histogram showing U87 cells stained with MBS1753716 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Wnt5b Antibody (MBS1753716,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit 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 U87 cells using anti-Wnt5b antibody (MBS1753716).Overlay histogram showing U87 cells stained with MBS1753716 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Wnt5b Antibody (MBS1753716,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Kanazawa, A., Tsukada, S., Sekine, A., Tsunoda, T., Takahashi, A., Kashiwagi, A., Tanaka, Y., Babazono, T., Matsuda, M., Kaku, K., Iwamoto, Y., Kawamori, R., Kikkawa, R., Nakamura, Y., Maeda, S. Association of the gene encoding wingless-type mammary tumor virus integration-site family member 5B (WNT5B) with type 2 diabetes. Am. J. Hum. Genet. 75: 832-843, 2004.
3. Saitoh, T., Katoh, M. Molecular cloning and characterization of human WNT5B on chromosome 12p13. 3 region. Int. J. Oncol. 19: 347-351, 2001.
NCBI and Uniprot Product Information
Observed: 465 kDa
NCBI Description
The WNT gene family consists of structurally related genes which encode secreted signaling proteins. These proteins have been implicated in oncogenesis and in several developmental processes, including regulation of cell fate and patterning during embryogenesis. This gene is a member of the WNT gene family. It encodes a protein which shows 94% and 80% amino acid identity to the mouse Wnt5b protein and the human WNT5A protein, respectively. Alternative splicing of this gene generates 2 transcript variants. [provided by RefSeq, Jul 2008]
Uniprot Description
WNT5B: Ligand for members of the frizzled family of seven transmembrane receptors. Probable developmental protein. May be a signaling molecule which affects the development of discrete regions of tissues. Is likely to signal over only few cell diameters. Interacts with PORCN. Belongs to the Wnt family.
Protein type: Secreted; Secreted, signal peptide
Chromosomal Location of Human Ortholog: 12p13.3
Cellular Component: proteinaceous extracellular matrix; extracellular space; cell surface; Golgi lumen; endoplasmic reticulum lumen; extracellular region; plasma membrane
Molecular Function: frizzled binding; receptor binding
Biological Process: fat cell differentiation; neuron differentiation; Wnt receptor signaling pathway; cell fate commitment; wound healing; chondrocyte differentiation; positive regulation of fat cell differentiation; positive regulation of cell migration
Research Articles on WNT5B
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Product Notes
The WNT5B wnt5b (Catalog #AAA1753716) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-Wnt5b Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's Wnt5b can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the WNT5B wnt5b 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 "Wnt5b, Polyclonal 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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