Western Blot (WB)
(Figure 1. Western blot analysis of NFAT5 using anti-NFAT5 antibody (MBS1753466).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 30ug of sample under reducing conditions.Lane 1: human THP-1 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-NFAT5 antigen affinity purified polyclonal antibody (Catalog # MBS1753466) 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 NFAT5 at approximately 170KD. The expected band size for NFAT5 is at 170KD. )
Rabbit anti-Human NFAT5 Polyclonal Antibody | anti-NFAT5 antibody
Anti-NFAT5 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 NFAT5 using anti-NFAT5 antibody (MBS1753466).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 30ug of sample under reducing conditions.Lane 1: human THP-1 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-NFAT5 antigen affinity purified polyclonal antibody (Catalog # MBS1753466) 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 NFAT5 at approximately 170KD. The expected band size for NFAT5 is at 170KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of NFAT5 using anti-NFAT5 antibody (MBS1753466).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 30ug of sample under reducing conditions.Lane 1: human THP-1 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-NFAT5 antigen affinity purified polyclonal antibody (Catalog # MBS1753466) 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 NFAT5 at approximately 170KD. The expected band size for NFAT5 is at 170KD. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of HepG2 cells using anti-NFAT5 antibody (MBS1753466).Overlay histogram showing HepG2 cells stained with MBS1753466 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NFAT5 Antibody (MBS1753466, 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 HepG2 cells using anti-NFAT5 antibody (MBS1753466).Overlay histogram showing HepG2 cells stained with MBS1753466 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NFAT5 Antibody (MBS1753466, 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. Lopez-Rodriguez, C., Antos, C. L., Shelton, J. M., Richardson, J. A., Lin, F., Novobrantseva, T. I., Bronson, R. T., Igarashi, P., Rao, A., Olson, E. N. Loss of NFAT5 results in renal atrophy and lack of tonicity-responsive gene expression. Proc. Nat. Acad. Sci. 101: 2392-2397, 2004.
3. Lopez-Rodriguez, C., Aramburu, J., Rakeman, A. S., Rao, A. NFAT5, a constitutively nuclear NFAT protein that does not cooperate with Fos and Jun. Proc. Nat. Acad. Sci. 96: 7214-7219, 1999.
NCBI and Uniprot Product Information
NCBI Description
The product of this gene is a member of the nuclear factors of activated T cells family of transcription factors. Proteins belonging to this family play a central role in inducible gene transcription during the immune response. This protein regulates gene expression induced by osmotic stress in mammalian cells. Unlike monomeric members of this protein family, this protein exists as a homodimer and forms stable dimers with DNA elements. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2008]
Uniprot Description
NFAT5: Plays a role in the inducible expression of genes. Regulates hypertonicity-induced cellular accumulation of osmolytes. Homodimer when bound to DNA, completely encircles its DNA target. Does not bind with Fos and Jun transcription factors. Highest levels in skeletal muscle, brain, heart and peripheral blood leukocytes. Also expressed in placenta, lung, liver, kidney, pancreas, spleen, thymus, prostate, testis, ovary, small intestine and colon. 4 isoforms of the human protein are produced by alternative splicing.
Protein type: Transcription factor
Chromosomal Location of Human Ortholog: 16q22.1
Cellular Component: cytoplasm; nucleus
Molecular Function: protein binding; transcription factor activity
Biological Process: transcription from RNA polymerase II promoter; cytokine production; positive regulation of transcription from RNA polymerase II promoter; excretion; signal transduction
Research Articles on NFAT5
Similar Products
Product Notes
The NFAT5 nfat5 (Catalog #AAA1753466) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-NFAT5 Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's NFAT5 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 NFAT5 nfat5 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 "NFAT5, 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
If you are ready to order, navigate to Shopping Cart and get ready to checkout.
