WB
Western blot analysis of NFATc3 on different lysates with Rabbit anti-NFATc3 antibody at 1/5,000 dilution. Lane 1: Jurkat cell lysate, Lane 2: Daudi cell lysate, Lane 3: HeLa cell lysate, Lane 4: Raji cell lysate, Lane 5: NIH/3T3 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: Lane 1-4: 20 seconds; Lane 5: 1 minute 33 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human thymus tissue with Rabbit anti-NFATc3 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-NFATc3 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-NFATc3 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of K-562 cells labeling NFATc3 with Rabbit anti-NFATc3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NFATc3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of K-562 cells labeling NFATc3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
NFATc3 was immunoprecipitated from 0.2 mg K-562 cell lysate with Rabbit anti-NFATc3 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-NFATc3 antibody at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input), Lane 2: Rabbit anti-NFATc3 antibody IP in K-562 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-NFATc3 antibody in K-562 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 20 seconds.| Product Name | NFATc3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human NFATc3 aa 1-450. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000 IHC:1:200 ICC/IF:1:100 FC:1:1000 IP: 1-2μg/sample |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NFATC3 |
|---|---|
| Gene Synonyms | NFAT4 NFATX NF-AT4c n339260 |
| Gene Full Name | nuclear factor of activated T cells 3 |
| Gene Summary | The product of this gene is a member of the nuclear factors of activated T cells DNA-binding transcription complex. This complex consists of at least two components: a preexisting cytosolic component that translocates to the nucleus upon T cell receptor (TCR) stimulation and an inducible nuclear component. Other members of this family participate to form this complex also. The product of this gene plays a role in the regulation of gene expression in T cells and immature thymocytes. Several transcript variants encoding distinct isoforms have been identified for this gene. [provided by RefSeq, Nov 2010] |
| Alternative Names | C80703 antibody cytoplasmic 3 antibody D8Ertd281e antibody NF AT4 antibody NF ATc3 antibody NF-AT4 antibody NF-ATc3 antibody NFAC3_HUMAN antibody NFAT4 antibody NFATc3 antibody C80703 antibody cytoplasmic 3 antibody D8Ertd281e antibody NF AT4 antibody NF ATc3 antibody NF-AT4 antibody NF-ATc3 antibody NFAC3_HUMAN antibody NFAT4 antibody NFATc3 antibody NFATx antibody Nuclear factor of activated T cells cytoplasmic 3 antibody nuclear factor of activated T cells cytoplasmic calcineurin dependent 3 antibody Nuclear factor of activated T-cells antibody T cell transcription factor NFAT4 antibody T-cell transcription factor NFAT4 antibody |
| Molecular Weight(MW) | 116 kDa |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of NFATc3 on different lysates with Rabbit anti-NFATc3 antibody at 1/5,000 dilution. Lane 1: Jurkat cell lysate, Lane 2: Daudi cell lysate, Lane 3: HeLa cell lysate, Lane 4: Raji cell lysate, Lane 5: NIH/3T3 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: Lane 1-4: 20 seconds; Lane 5: 1 minute 33 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human thymus tissue with Rabbit anti-NFATc3 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-NFATc3 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-NFATc3 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of K-562 cells labeling NFATc3 with Rabbit anti-NFATc3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NFATc3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of K-562 cells labeling NFATc3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
NFATc3 was immunoprecipitated from 0.2 mg K-562 cell lysate with Rabbit anti-NFATc3 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-NFATc3 antibody at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input), Lane 2: Rabbit anti-NFATc3 antibody IP in K-562 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-NFATc3 antibody in K-562 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 20 seconds.| Application Notes | WB:1:5000 IHC:1:200 ICC/IF:1:100 FC:1:1000 IP: 1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
