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USP15 Recombinant Rabbit Monoclonal Antibody [PSH15-94]

Usd: 385

Specification

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Catalog# HA723799

USP15 Recombinant Rabbit Monoclonal Antibody [PSH15-94]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

USP15 Recombinant Rabbit Monoclonal Antibody [PSH15-94]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human USP15 aa 500-795.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 112 kDa

Positive Control

HeLa cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, COS-1 cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, mouse kidney tissue, mouse liver tissue, rat kidney tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

PSH15-94

Reactivity Data

WB IHC-P IP
Human
Tested Tested
Not Recommended
Tested Tested
Mouse
Tested Tested
Tested Tested
Rat
Tested Tested
Tested Tested
Green Monkey
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:20,000

  • IHC-P

  • 1:200-1:1,000

  • IP

  • 1-2μg/sample

Target

Function

USP15 is a hydrolase that removes conjugated ubiquitin from target proteins and regulates pathways like TGF-beta receptor signaling and NF-kappa-B. It acts as a key regulator of the TGF-beta receptor signaling pathway, possibly by promoting deubiquitination of R-SMADs (SMAD1, SMAD2 and/or SMAD3), which helps activate TGF-beta target genes. USP15 can also deubiquitinate monoubiquitinated substrates and various polyubiquitin chains ('Lys-27', 'Lys-48', 'Lys-63'). Additionally, it may regulate gene expression and DNA repair through histone H2B deubiquitination. USP15 inhibits mitophagy by counteracting parkin (PRKN), hydrolyzing specific polyubiquitin chains on target proteins like MFN2. It also associates with the COP9 signalosome complex (CSN) to regulate different pathways, including NF-kappa-B via deubiquitination of NFKBIA and substrates bound to VCP. USP15 is involved in endosome organization by deubiquitinating SQSTM1 and acts as a negative regulator of antifungal immunity by deubiquitinating CARD9.

Background References

1. Huangfu L et al. The deubiquitinase USP15 drives malignant progression of gastric cancer through glucose metabolism remodeling. J Exp Clin Cancer Res. 2024 Aug

2. Kim MJ et al. USP15 negatively regulates lung cancer progression through the TRAF6-BECN1 signaling axis for autophagy induction. Cell Death Dis. 2022 Apr

Subcellular Location

Cytoplasm, Nucleus, Mitochondrion.

UNIPROT

SwissProt: Q9Y4E8 Human

SwissProt: Q8R5H1 Mouse

SwissProt: Q9R085 Rat

Synonyms

Deubiquitinating enzyme 15 antibody

Deubiquitinating enzyme antibody

EC 3 1 2 15 antibody

KIAA0529 antibody

MGC131982 antibody

MGC149838 antibody

MGC74854 antibody

Ubiquitin Carboxy terminal Hydrolase 15 antibody

Ubiquitin carboxyl-terminal hydrolase 15 antibody

Ubiquitin specific peptidase 15 antibody

Expand

Images

  • Western blot analysis of USP15 on different lysates with Rabbit anti-USP15 antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/20,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: K-562 cell lysate (20 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 4: PC-12 cell lysate (20 µg/Lane)<br />Lane 5: COS-1 cell lysate (20 µg/Lane)<br />Lane 6: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 7: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 112 kDa<br />Observed band size: 112 kDa<br /><br />Exposure time: 4 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of USP15 on different lysates with Rabbit anti-USP15 antibody (HA723799) at 1/20,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: K-562 cell lysate (20 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 4: PC-12 cell lysate (20 µg/Lane)
    Lane 5: COS-1 cell lysate (20 µg/Lane)
    Lane 6: Mouse liver tissue lysate (40 µg/Lane)
    Lane 7: Rat liver tissue lysate (40 µg/Lane)

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

    Exposure time: 4 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 (HA723799) at 1/20,000 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.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-USP15 antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/1,000 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.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-USP15 antibody (HA723799) at 1/1,000 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 (HA723799) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-USP15 antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-USP15 antibody (HA723799) 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 (HA723799) 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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-USP15 antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/1,000 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.

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-USP15 antibody (HA723799) at 1/1,000 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 (HA723799) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-USP15 antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a>) at 1/1,000 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.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-USP15 antibody (HA723799) at 1/1,000 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 (HA723799) at 1/1,000 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.

  • USP15 was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a> 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.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA723799" style="font-weight: bold;text-decoration: underline;">HA723799</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 25 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    USP15 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA723799 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723799 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: HA723799 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA723799 in HeLa cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 25 seconds; ECL: K1801

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