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, Monkey
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: JJ08-40
Form: Liquid
Storage condition: 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*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: PC-12 cell lysate, COS-1 cell lysate, Mouse brain tissue lysate, Mouse kidney tissue lysate, Rat brain tissue lysate, Rat liver tissue lysate, 293T cell lysate, SH-SY5Y cell lysate, PC-12, Hela, NIH/3T3, human brain tissue, mouse brain tissue, mouse kidney tissue, rat brain tissue, rat liver tissue.
Subcellular location: Nucleus, Cytoplasm, Melanosome.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:5,000
1:50-1:200
1:50-1:200
1:200
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
ET1701-1_1.jpg Fig1: Western blot analysis of 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-14-3-3 epsilon KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 29 kDa
Observed band size: 29 kDa

Exposure time: 20 seconds; ECL: K1801;

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 (ET1701-1) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1701-1_2.jpg Fig2: Western blot analysis of 14-3-3 epsilon on different lysates with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) at 1/5,000 dilution.

Lane 1: PC-12 cell lysate (20 µg/Lane)
Lane 2: COS-1 cell lysate (20 µg/Lane)
Lane 3: Mouse brain tissue lysate (40 µg/Lane)
Lane 4: Mouse kidney tissue lysate (40 µg/Lane)
Lane 5: Rat brain tissue lysate (40 µg/Lane)
Lane 6: Rat liver tissue lysate (40 µg/Lane)

Predicted band size: 29 kDa
Observed band size: 29 kDa

Exposure time: 10 seconds; ECL: K1801;

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 (ET1701-1) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1701-1_3.jpg Fig3: 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: 29 kDa
Observed band size: 29 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.
ET1701-1_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling 14-3-3 epsilon with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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-14-3-3 epsilon antibody (ET1701-1) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1701-1_5.jpg Fig5: 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).
ET1701-1_6.jpg Fig6: 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).
ET1701-1_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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.
ET1701-1_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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.
ET1701-1_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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.
ET1701-1_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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.
ET1701-1_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-14-3-3 epsilon antibody (ET1701-1) 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 (ET1701-1) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.