Application
Immunoprecipitation of NFAT5frpm U2OS cells. The antigen-antibody complex was formed by incubating 500μg whole cell lysate with 3μg of NFAT5 antibody overnight on a rocking platform at 4°C. The immune-complex was captured on 50μl Protein A/G Agarose. Captured immune-complexes were washed and eluted. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% Milk/TBS-0.1%Tween for at least 1 hour. Membranes were washed in TBS-0.1%Tween 20 and probed with a proper secondary antibody. Membranes were washed and chemiluminescent detection performed.Application
Immunofluorescent analysis of NFAT5 in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 1% BSA for 15 minutes at room temperature. Cells were probed with NFAT5 antibody at a dilution of 1:100 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with a proper secondary antibody. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.Application
Western blot analysis of NFAT5 in 25ug of various whole cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with NFAT5 antibody at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a HRP-conjugated secondary antibody. Membranes were washed and chemiluminescent detection performed.| Product Name | NFAT5 antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to NFAT5The 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] |
| Immunogen | Synthetic peptide corresponding to residues C D(1439) L L V S L Q N Q G N N L T G S F(1455) of human NFAT5. |
| Conjugation | Agarose |
| Modification | Acetylated |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPGSAICC/IFIHCIHC-PIPWB |
| WB:1:500~1:1000 IP:1:100~1:500 ICC/IF:1:100~1:500 IHC:1:50~1:100 |
|
| Species Reactivity | HumanMousePigRat |
| Concentration | 1mg/ml (Please refer to the vial label for the specific concentration) |
| Purification | Affinity purified |
| Gene Synonyms | NF-AT5 NFATL1 nuclear factor of activated T-cells 5 tonicity-responsive OREBP TONEBP NFATZ |
|---|---|
| Target | NFAT5 |
Application
Immunoprecipitation of NFAT5frpm U2OS cells. The antigen-antibody complex was formed by incubating 500μg whole cell lysate with 3μg of NFAT5 antibody overnight on a rocking platform at 4°C. The immune-complex was captured on 50μl Protein A/G Agarose. Captured immune-complexes were washed and eluted. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% Milk/TBS-0.1%Tween for at least 1 hour. Membranes were washed in TBS-0.1%Tween 20 and probed with a proper secondary antibody. Membranes were washed and chemiluminescent detection performed.
Application
Immunofluorescent analysis of NFAT5 in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 1% BSA for 15 minutes at room temperature. Cells were probed with NFAT5 antibody at a dilution of 1:100 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with a proper secondary antibody. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.
Application
Western blot analysis of NFAT5 in 25ug of various whole cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with NFAT5 antibody at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a HRP-conjugated secondary antibody. Membranes were washed and chemiluminescent detection performed.| Application Notes | WB:1:500~1:1000 IP:1:100~1:500 ICC/IF:1:100~1:500 IHC:1:50~1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | For short-term storage, store at 4° C. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. |
