ET1701-1

14-3-3 epsilon Recombinant Rabbit Monoclonal Antibody [JJ08-40]

cat.: ET1701-1

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JJ08-40

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: 29 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human 14-3-3 epsilon aa 51-100.
Positive control:	293T cell lysate, SH-SY5Y cell lysate, Hela, SH-SY5Y, NIH/3T3, mouse placenta tissue, human colon carcinoma tissue.
Subcellular location:	Nucleus, Cytoplasm, Melanosome.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:1,000-1:2,000 1:50-1:200 1:50-1:200 1:50-1:200 1:10-1:100
Uniprot #:	SwissProt: P62258 Human \| P62259 Mouse \| P62260 Rat
Alternative names:	14 3 3 E 14 3 3 epsilon 14 3 3E 14-3-3 protein epsilon 14-3-3E 1433E_HUMAN Epididymis luminal protein 2 FLJ45465 FLJ53559 HEL2 KCIP 1 KCIP1 MDCR MDS Mitochondrial import stimulation factor L subunit Protein kinase C inhibitor protein1 Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein, epsilon Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein, epsilon polypeptide Tyrosine 3/tryptophan 5 monooxygenase activation protein epsilon polypeptide YWHAE

Images

	Fig1: Western blot analysis of 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) at 1/500 dilution. Lane 1: 293T cell lysate Lane 2: SH-SY5Y cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 28 kDa Observed band size: 28 kDa Exposure time: 30 seconds; 12% 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 (ET1701-1) 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 14-3-3 epsilon in Hela cells (green). 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 (ET1701-1, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 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 14-3-3 epsilon in SH-SY5Y cells (green). 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 (ET1701-1, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 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 14-3-3 epsilon in NIH/3T3 cells (green). 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 (ET1701-1, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 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 mouse placenta tissue using anti-14-3-3 epsilon 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 (ET1701-1, 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 colon carcinoma tissue using anti-14-3-3 epsilon 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 (ET1701-1, 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: Flow cytometric analysis of 14-3-3 epsilon was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-1, 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 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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