USP14 Recombinant Rabbit Monoclonal Antibody [JU30-49]
cat.: ET1706-37
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Zebrafish
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JU30-49
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: 56 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human USP14 aa 351-494 / 494.
Positive control: K562 cell lysate, Hela cell lysate, zebrafish lysates, A431, HUVEC, LOVO, human colon carcinoma tissue, human kidney tissue, mouse prostate tissue, rat epididymis tissue, Jurkat.
Subcellular location: Cytoplasm. Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:2,000
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P54578 Human
Alternative names: Deubiquitinating enzyme 14 TGT tRNA guanine transglycosylase 60 kD subunit tRNA guanine transglycosylase Ubiquitin carboxyl terminal hydrolase 14 Ubiquitin carboxyl-terminal hydrolase 14 Ubiquitin specific peptidase 14 Ubiquitin specific processing protease 14 Ubiquitin specific protease 14 Ubiquitin thiolesterase 14 Ubiquitin-specific-processing protease 14 UBP14_HUMAN USP 14 USP14
Images
ET1706-37_1.jpg Fig1: Western blot analysis of USP14 on different lysates with Rabbit anti-USP14 antibody (ET1706-37) at 1/1,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si USP14 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 60 kDa

Exposure time: 12 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 (ET1706-37) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1706-37_2.jpg Fig2: Western blot analysis of USP14 on different lysates with Rabbit anti-USP14 antibody (ET1706-37) at 1/500 dilution.

Lane 1: K562 cell lysate
Lane 2: Hela cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 60 kDa

Exposure time: 2 minutes;

10% 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 (ET1706-37) 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 200,000 dilution was used for 1 hour at room temperature.
ET1706-37_3.jpg Fig3: Western blot analysis of USP14 on zebrafish lysates with Rabbit anti-USP14 antibody (ET1706-37) at 1/500 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 60 kDa

Exposure time: 2 minutes;

10% 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 (ET1706-37) 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 1:200,000 dilution was used for 1 hour at room temperature.
ET1706-37_4.jpg Fig4: Immunocytochemistry analysis of A431 cells labeling USP14 with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-37_5.jpg Fig5: Immunocytochemistry analysis of HUVEC cells labeling USP14 with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-37_6.jpg Fig6: Immunocytochemistry analysis of LOVO cells labeling USP14 with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-37_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 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 (ET1706-37) at 1/50 dilution for 0.5 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.
ET1706-37_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 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 (ET1706-37) at 1/50 dilution for 0.5 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.
ET1706-37_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse prostate tissue with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 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 (ET1706-37) at 1/50 dilution for 0.5 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.
ET1706-37_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat epididymis tissue with Rabbit anti-USP14 antibody (ET1706-37) at 1/50 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 (ET1706-37) at 1/50 dilution for 0.5 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.
ET1706-37_11.jpg Fig11: Flow cytometric analysis of USP14 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1706-37, 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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