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Ubiquitin Specific Peptidase 18 Peptide 1

Antibody: CPTC-USP18-1
Background

Catalog Number:

CPTC-USP18-1

Target Antigen:

Ubiquitin Specific Peptidase 18 Peptide 1

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

02/09/2024

Antigen Recognition(s):

Peptide, Endogenous

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Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry

CPTC-USP18-1 Affinity and Kinetics (Bio-Layer Interferometry) 

Result: No Binding

The affinity and binding kinetics of CPTC-USP18-1 antibody and positive control BSA-conjugated human UBP43 (USP18) peptide (amino acids 233 - 250) and negative control BSA-conjugated human UBP43 (USP18) peptide (amino acids 199 - 215) were measured using biolayer interferometry. In separate assays, each respective BSA-conjugated peptide was covalently immobilized onto AR2G biosensors using standard amine coupling.   CPTC-USP18-1 antibody was used as analyte and was titrated at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM. Buffer only and biosensors immobilized without BSA-conjugated peptide were used as references for background subtraction. All data was analyzed globally using a 1:2 bivalent fitting model.  The positive control peptide showed non-specific binding > 20% and the negative control peptide showed no binding.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-USP18-1 and BSA-conjugated human UBP43 (USP18) (amino acids 233-250) peptide were measured using surface plasmon resonance. BSA conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-USP18-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM and 1.0 nM was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model. Click image to enlarge

CPTC-USP18-1 Affinity and Kinetics (Surface Plasmon Resonance) 

Result: High Binding

Affinity and binding kinetics of CPTC-USP18-1 and BSA-conjugated human UBP43 (USP18) (amino acids 233-250) peptide were measured using surface plasmon resonance. BSA conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-USP18-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM and 1.0 nM was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  Flow Cytometry

CPTC-USP18-1 (Flow Cytometry)

Result: Negative

Flow cytometric analysis of USP18 expression in A549-USP18 and A549-Empty Vector cells or H1299-USP18 and H1299 Empty Vector cells using CPTC-USP18-1 rabbit antibody. A549-USP18 cells (solid green) or A549-Empty Vector cells (dashed green) were permeabilized and fixed and then stained with CPTC-USP18-1 antibody. H1299-USP18 cells (solid blue) or H1299 Empty Vector cells (dashed blue) were permeabilized and fixed and then stained with CPTC-USP18-1 antibody. An APC conjugated goat anti-rabbit IgG (H+L) was used as a secondary antibody. All data were analyzed using FlowJo.


Characterization SOP Files

  IHC Tissue
Click to enlarge image Tissue Micro-Array (TMA) core of showing breast cancer showing cytoplasmic and membranous staining using Antibody CPTC-USP18-1. Titer: 1:1000 Click image to enlarge

CPTC-USP18-1 IHC Tissue

Result: Positive

Tissue Micro-Array (TMA) core of showing breast cancer showing cytoplasmic and membranous staining using Antibody CPTC-USP18-1. Titer: 1:1000


Characterization SOP Files

  Immunofluorescence

CPTC-USP18-1 Immunofluorescence

Result: Negative

Immunofluorescence staining of human cell lines A549 and H1299 transfected with Green Fluorescent Protein (GFP) and Empty Vector (EV) using CPTC-USP18-1 as primary antibody. USP18 protein expression shows no localization.


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA using CPTC-USP18-1 as primary rabbit antibody against BSA-conjugated human UBP43 (USP18) peptide (amino acids 233-250) (A.) and BSA-conjugated human UBP43 (USP18) peptide (amino acids 199-215) (B.) coated on the plate and detected using the goat anti-rabbit antibody and TMB.  Click image to enlarge

CPTC-USP18-1 Indirect ELISA

Result: High Binding

Indirect ELISA using CPTC-USP18-1 as primary rabbit antibody against BSA-conjugated human UBP43 (USP18) peptide (amino acids 233-250) (A.) and BSA-conjugated human UBP43 (USP18) peptide (amino acids 199-215) (B.) coated on the plate and detected using the goat anti-rabbit antibody and TMB. 


Characterization SOP Files

  NCI 60 Protein Array

CPTC-USP18-1 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate

CPTC-USP18-1 Single Cell Western Blot (Cell Lysate)

Result: Negative

Single cell western blot using CPTC-USP18-1 as a primary antibody against human cell lines A549 and H1299 transfected with Green Fluorescent Protein (GFP) and Empty Vector (EV) cell lysates. USP18 protein expression was not detected.


Characterization SOP Files

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Antigen: Ubiquitin Specific Peptidase 18 Peptide 1
Background

NCI Identification Number:

00534

Antigen Name:

Ubiquitin Specific Peptidase 18 Peptide 1

CPTC Name:

CPTC-USP18 Peptide 1

Aliases:

Ubiquitin Specific Peptidase 18; Ubl Carboxyl-Terminal Hydrolase 18; ISG15-Specific-Processing Protease; Ubiquitin Specific Protease 18; 43 KDa ISG15-Specific Protease; Ubl Thioesterase 18; ISG43; Ubl Thiolesterase 1; EC 3.4.19.-; EC 3.1.2.15; PTORCH2; HUBP43; UBP43

Function:

The protein encoded by this gene belongs to the ubiquitin-specific proteases (UBP) family of enzymes that cleave ubiquitin from ubiquitinated protein substrates. It is highly expressed in liver and thymus, and is localized to the nucleus. This protein efficiently cleaves only ISG15 (a ubiquitin-like protein) fusions, and deletion of this gene in mice results in a massive increase of ISG15 conjugates in tissues, indicating that this protein is a major ISG15-specific protease. Mice lacking this gene are also hypersensitive to interferon, suggesting a function of this protein in downregulating interferon responses, independent of its isopeptidase activity towards ISG15. USP18 (Ubiquitin Specific Peptidase 18) is a Protein Coding gene. Diseases associated with USP18 include Pseudo-Torch Syndrome 2 and Torch Syndrome. Among its related pathways are MyD88 dependent cascade initiated on endosome and Deubiquitination. Gene Ontology (GO) annotations related to this gene include cysteine-type endopeptidase activity and cysteine-type deubiquitinase activity. An important paralog of this gene is USP9X.Involved in the negative regulation of the inflammatory response triggered by type I interferon (PubMed:28165510, 27325888). Upon recruitment by STAT2 to the type I IFN receptor subunit IFNAR2 interferes with the assembly of the ternary interferon-IFNAR1-IFNAR2 complex and acts as a negative regulator of the type I IFN signaling pathway (PubMed:28165510). Also regulates protein ISGylation. Can efficiently cleave only ISG15 fusions including native ISG15 conjugates linked via isopeptide bonds. Necessary to maintain a critical cellular balance of ISG15-conjugated proteins in both healthy and stressed organisms (PubMed:11788588). ( UBP18_HUMAN,Q9UMW8 )[Isoform 2]: Has enzymatic activity similar to isoform 1 and interferes with type I interferon signaling. Major deISGylation enzyme for nuclear proteins (PubMed:22170061). ( UBP18_HUMAN,Q9UMW8 )

Chromosomal Localization:

22q11.2

Accession Number:

XP_006724137.1

UniProt Accession Number:

Q9UMW8

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

CGKKTRGKQVLKLTHLPQ

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

1980

Last Updated:

10/13/2022

Links

Characterization Data

SOPs

No SOPs available.

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