OTUB1 Recombinant Rabbit Monoclonal Antibody [PSH18-50]
cat.: HA751674
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IHC-P, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: PSH18-50
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 31 kDa
Isotype: IgG
Immunogen: Recombinant protein within human OTUB1 aa 1-271.
Positive control: HEK-293 cell lysate, HeLa cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, COS-1 cell lysate, Mouse brian (C57BL/6) tissue lysate, Rat brain tissue lysate, human colon tissue, human placenta tissue, mouse colon tissue, mouse placenta tissue, rat colon tissue, rat placenta tissue, HeLa, NIH/3T3.
Subcellular location: Cytoplasm, nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
  IP
1:5,000-1:25,000
1:200
1:100
1:1,000
1-2μg/sample
Uniprot #: SwissProt: Q96FW1 Human | Q7TQI3 Mouse | B2RYG6 Rat
Alternative names: OTB1 OTU1 HSPC263 OTUB1 Ubiquitin thioesterase OTUB1 Deubiquitinating enzyme OTUB1 OTU domain-containing ubiquitin aldehyde-binding protein 1 Otubain-1 Ubiquitin-specific-processing protease OTUB1 hOTU1
Images
HA751674_1.jpg Fig1: Western blot analysis of OTUB1 on different lysates with Rabbit anti-OTUB1 antibody (HA751674) at 1/5,000 dilution.

Lane 1: HEK-293 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: MCF7 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: COS-1 cell lysate (20 µg/Lane)
Lane 6: Mouse brian (C57BL/6) tissue lysate (30 µg/Lane)
Lane 7: Rat brain tissue lysate (30 µg/Lane)

Predicted band size: 31 kDa
Observed band size: 33 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 (HA751674) 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.
HA751674_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-OTUB1 antibody (HA751674) 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 (HA751674) 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.
HA751674_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-OTUB1 antibody (HA751674) 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 (HA751674) 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.
HA751674_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-OTUB1 antibody (HA751674) 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 (HA751674) 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.
HA751674_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse placenta tissue with Rabbit anti-OTUB1 antibody (HA751674) 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 (HA751674) 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.
HA751674_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-OTUB1 antibody (HA751674) 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 (HA751674) 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.
HA751674_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat placenta tissue with Rabbit anti-OTUB1 antibody (HA751674) 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 (HA751674) 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.
HA751674_8.jpg Fig8: Immunocytochemistry analysis of HeLa cells labeling OTUB1 with Rabbit anti-OTUB1 antibody (HA751674) 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-OTUB1 antibody (HA751674) 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.
HA751674_9.jpg Fig9: Immunocytochemistry analysis of NIH/3T3 cells labeling OTUB1 with Rabbit anti-OTUB1 antibody (HA751674) 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-OTUB1 antibody (HA751674) 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.
HA751674_10.jpg Fig10: Flow cytometric analysis of HeLa cells labeling OTUB1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751674, 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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA751674_11.jpg Fig11: Flow cytometric analysis of NIH/3T3 cells labeling OTUB1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751674, 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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA751674_12.jpg Fig12: OTUB1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751674 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751674 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: HeLa cell lysate (input)
Lane 2: HA751674 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751674 in HeLa cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 10 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.