ET1704-14

NFAT1 Recombinant Rabbit Monoclonal Antibody [JA11-08]

cat.: ET1704-14

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JA11-08

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 100 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human NFAT1 aa 21-70 / 925.
Positive control:	MCF-7, Hela, SHG-44, human spleen tissue, human tonsil tissue, human colon carcinoma tissue, Jurkat, MCF-7 cell lysate, Daudi cell lysate.
Subcellular location:	Cytoplasm. Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:500 1:50-1:200 1:50-1:200 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: Q13469 Human
Alternative names:	AI607462 cytoplasmic 2 KIAA0611 NF ATc2 NF ATp NF-ATc2 NF-ATp NFAC2_HUMAN NFAT 1 NFAT pre existing subunit NFAT pre-existing subunit NFAT transcription complex, preexisting component NFAT1 NFAT1-D NFATc2 NFATp Nuclear factor of activated T cells cytoplasmic 2 Nuclear factor of activated T cells cytoplasmic calcineurin dependent 2 Nuclear factor of activated T cells pre-existing component Nuclear factor of activated T cells, preexisting component Nuclear factor of activated T-cells Preexisting nuclear factor of activated T cells 2 T cell transcription factor NFAT 1 T-cell transcription factor NFAT1

Images

	Fig1: Western blot analysis of NFAT1 on different lysates with Rabbit anti-NFAT1 antibody (ET1704-14) at 1/500 dilution. Lane 1: MCF-7 cell lysate Lane 2: Daudi cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 100 kDa Observed band size: 130 kDa Exposure time: 2 minutes; 6% 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 (ET1704-14) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
	Fig2: ICC staining of NFAT1 in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig3: ICC staining of NFAT1 in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: ICC staining of NFAT1 in SHG-44 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

	Fig5: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-NFAT1 antibody. 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 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-14, 1/50) for 30 minutes 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-NFAT1 antibody. 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 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-14, 1/50) for 30 minutes 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-NFAT1 antibody. 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 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-14, 1/50) for 30 minutes 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.

Fig8: Flow cytometric analysis of NFAT1 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-14, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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