Western Blot (WB)
(Figure 1. Western blot analysis of DR3/Tnfrsf25 using anti-DR3/Tnfrsf25 antibody (MBS1753579).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: mouse SP2/0 whole cell lysatesLane 2: mouse RAW264. 7 whole cell lysatesLane 3: mouse NIH/3T3 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-DR3/Tnfrsf25 antigen affinity purified polyclonal antibody (Catalog # MBS1753579) 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 1:5000 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 DR3/Tnfrsf25 at approximately 60KD. The expected band size for DR3/Tnfrsf25 is at 60KD. )
Rabbit anti-Mouse, Rat DR3/Tnfrsf25 Polyclonal Antibody | anti-Tnfrsf25 antibody
Anti-DR3/Tnfrsf25 Antibody
FC/FACS/FCM: 1-3ug/1x106 cells|Rat|
Direct ELISA: 0.1-0.5ug/ml|Rat|
Western Blot (WB)
(Figure 1. Western blot analysis of DR3/Tnfrsf25 using anti-DR3/Tnfrsf25 antibody (MBS1753579).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: mouse SP2/0 whole cell lysatesLane 2: mouse RAW264. 7 whole cell lysatesLane 3: mouse NIH/3T3 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-DR3/Tnfrsf25 antigen affinity purified polyclonal antibody (Catalog # MBS1753579) 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 1:5000 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 DR3/Tnfrsf25 at approximately 60KD. The expected band size for DR3/Tnfrsf25 is at 60KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of DR3/Tnfrsf25 using anti-DR3/Tnfrsf25 antibody (MBS1753579).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: mouse SP2/0 whole cell lysatesLane 2: mouse RAW264. 7 whole cell lysatesLane 3: mouse NIH/3T3 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-DR3/Tnfrsf25 antigen affinity purified polyclonal antibody (Catalog # MBS1753579) 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 1:5000 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 DR3/Tnfrsf25 at approximately 60KD. The expected band size for DR3/Tnfrsf25 is at 60KD. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of NRK cells using anti-DR3/Tnfrsf25 antibody (MBS1753579).Overlay histogram showing NRK cells stained with MBS1753579 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DR3/Tnfrsf25 Antibody (MBS1753579, 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 NRK cells using anti-DR3/Tnfrsf25 antibody (MBS1753579).Overlay histogram showing NRK cells stained with MBS1753579 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DR3/Tnfrsf25 Antibody (MBS1753579, 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. Fang, L., Adkins, B., Deyev, V., Podack, E. R. Essential role of TNF receptor superfamily 25 (TNFRSF25) in the development of allergic lung inflammation. J. Exp. Med. 205: 1037-1048, 2008.
3. Marsters, S. A., Sheridan, J. P., Donahue, C. J., Pitti, R. M., Gray, C. L., Goddard, A. D., Bauer, K. D., Ashkenazi, A. Apo-3, a new member of the tumor necrosis factor receptor family, contains a death domain and activates apoptosis and NF-kappa-B. Curr. Biol. 6: 1669-1676, 1996.
NCBI and Uniprot Product Information
Uniprot Description
TNFRSF25: Receptor for TNFSF12/APO3L/TWEAK. Interacts directly with the adapter TRADD. Mediates activation of NF-kappa-B and induces apoptosis. May play a role in regulating lymphocyte homeostasis. Homodimer. Interacts strongly via the death domains with TNFRSF1 and TRADD to activate at least two distinct signaling cascades, apoptosis and NF-kappa-B signaling. Interacts with BAG4. Abundantly expressed in thymocytes and lymphocytes. Detected in lymphocyte-rich tissues such as thymus, colon, intestine, and spleen. Also found in the prostate. 12 isoforms of the human protein are produced by alternative splicing.
Protein type: Apoptosis; Membrane protein, integral; Receptor, misc.
Chromosomal Location of Human Ortholog: 5q36
Cellular Component: integral component of plasma membrane; plasma membrane
Molecular Function: tumor necrosis factor-activated receptor activity
Biological Process: immune response; inflammatory response; multicellular organism development; regulation of apoptosis; regulation of cell proliferation; response to lipopolysaccharide; tumor necrosis factor-mediated signaling pathway
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Product Notes
The Tnfrsf25 tnfrsf25 (Catalog #AAA1753579) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-DR3/Tnfrsf25 Antibody reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's DR3/Tnfrsf25 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|Mouse, Rat| FC/FACS/FCM: 1-3ug/1x106 cells|Rat| Direct ELISA: 0.1-0.5ug/ml|Rat|. Researchers should empirically determine the suitability of the Tnfrsf25 tnfrsf25 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 "DR3/Tnfrsf25, 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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