HA500497

USP15 Rabbit Polyclonal Antibody

cat.: HA500497

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Monkey
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Polyclonal

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:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 112 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human USP15 aa 500-795.
Positive control:	HeLa cell lysate, A549 cell lysate, 293T cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, COS-1 cell lysate, HeLa, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location:	Cytoplasm, Nucleus, Mitochondrion.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:5,000 1:200 1:200 1:1,000
Uniprot #:	SwissProt: Q9Y4E8 Human \| Q8R5H1 Mouse \| Q9R085 Rat
Alternative names:	Deubiquitinating enzyme 15 Deubiquitinating enzyme EC 3 1 2 15 KIAA0529 MGC131982 MGC149838 MGC74854 Ubiquitin Carboxy terminal Hydrolase 15 Ubiquitin carboxyl-terminal hydrolase 15 Ubiquitin specific peptidase 15 Ubiquitin specific processing protease 15 Ubiquitin Specific Protease 15 Ubiquitin thioesterase 15 Ubiquitin thiolesterase 15 Ubiquitin-specific-processing protease 15 UBP 15 UBP15 UBP15_HUMAN Unph 2 UNPH 4 Unph-2 Unph2 Unph4 USP 15 Usp15

Images

Fig1: Western blot analysis of USP15 on different lysates with Rabbit anti-USP15 antibody (HA500497) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: 293T cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: PC-12 cell lysate
Lane 6: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 112 kDa
Observed band size: 112 kDa

Exposure time: 8 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 (HA500497) 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.

Fig2: Immunocytochemistry analysis of HeLa cells labeling USP15 with Rabbit anti-USP15 antibody (HA500497) at 1/200 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-USP15 antibody (HA500497) at 1/200 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-USP15 antibody (HA500497) 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 (HA500497) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-USP15 antibody (HA500497) 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 (HA500497) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-USP15 antibody (HA500497) 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 (HA500497) 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.

Fig6: Flow cytometric analysis of HeLa cells labeling USP15.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500497, 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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.