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Notum, Palmitoleoyl-Protein Carboxylesterase

Antibody: CPTC-NOTUM-1
Background

Catalog Number:

CPTC-NOTUM-1

Target Antigen:

Notum, Palmitoleoyl-Protein Carboxylesterase

Isotype:

IgG1

Species:

Mouse Monoclonal Antibody

Last Updated:

08/16/2023

Antigen Recognition(s):

Recombinant Full-length

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

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

Result: No Binding

Affinity and binding kinetics of CPTC-NOTUM-1 and human Notum recombinant protein were measured using biolayer interferometry. Recombinant protein was immobilized onto AR2G biosensors using amine coupling. CPTC-NOTUM-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without peptide were used as references for background subtraction. Binding data was double referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  Affinity Measurement by SPR

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

Result: No Binding

Affinity and binding kinetics of CPTC-NOTUM-1 and human Notum recombinant protein was measured using surface plasmon resonance. Recombinant protein was amine coupled onto a Series S Protein CM5 biosensor chip. CPTC-NOTUM-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM was used as analyte. Binding data were double referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  IHC Tissue

CPTC-NOTUM-1 IHC Tissue

Result: Negative

This antibody is not suitable for use in an Immunohistochemistry format as described in SOP M-106.


Characterization SOP Files

  IP

CPTC-NOTUM-1 Immunoprecipitation

Result: Negative

Immunoprecipitation using CPTC-NOTUM-1 as Capture antibody to probe the whole lysate of SW620 cells and over-expressed lysate. The eluates were tested in WB, using CPTC-NOTUM-3 as detection antibody. The antibody is not able to precipitate the target protein in the tested lysate.


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-1 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antibody. The antibody recognized the target NOTUM. Click image to enlarge

CPTC-NOTUM-1 automated WB (over-expressed lysate)

Result: Positive

Automated Western Blot using antibody CPTC-NOTUM-1 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antibody. The antibody recognized the target NOTUM.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-1 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody did not recognize the target NOTUM in the tested cell line lysates. Click image to enlarge

CPTC-NOTUM-1 automated WB (lysate)

Result: Negative

Automated Western Blot using antibody CPTC-NOTUM-1 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody did not recognize the target NOTUM in the tested cell line lysates.


Characterization SOP Files

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Antibody: CPTC-NOTUM-2
Background

Catalog Number:

CPTC-NOTUM-2

Target Antigen:

Notum, Palmitoleoyl-Protein Carboxylesterase

Isotype:

IgG1

Species:

Mouse Monoclonal Antibody

Last Updated:

08/16/2023

Antigen Recognition(s):

Recombinant Full-length, Endogenous

Characterization Data [Compare Characterization Data]
  Affinity Measurement by SPR

CPTC-NOTUM-2 Affinity and Kinetics (Surface Plasmon Resonance)

Result: No Binding

Affinity and binding kinetics of CPTC-NOTUM-2 and human Notum recombinant protein was measured using surface plasmon resonance. Recombinant protein was amine coupled onto a Series S Protein CM5 biosensor chip. CPTC-NOTUM-2 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM was used as analyte. Binding data were double referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  IHC Tissue

CPTC-NOTUM-2 IHC Tissue

Result: Negative

This antibody is not suitable for use in an Immunohistochemistry format as described in SOP M-106.


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-2 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antobody. The antibody recognized the target NOTUM. Click image to enlarge

CPTC-NOTUM-2 automated WB (over-expressed lysate)

Result: Positive

Automated Western Blot using antibody CPTC-NOTUM-2 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antobody. The antibody recognized the target NOTUM.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-2 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody presumebly recognized the target NOTUM in the tested cell line lysates. Click image to enlarge

CPTC-NOTUM-2 automated WB (lysate)

Result: Presumed Positive (with additional bands)

Automated Western Blot using antibody CPTC-NOTUM-2 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody presumebly recognized the target NOTUM in the tested cell line lysates.


Characterization SOP Files

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Antibody: CPTC-NOTUM-3
Background

Catalog Number:

CPTC-NOTUM-3

Target Antigen:

Notum, Palmitoleoyl-Protein Carboxylesterase

Isotype:

IgG1

Species:

Mouse Monoclonal Antibody

Last Updated:

08/16/2023

Antigen Recognition(s):

Recombinant Full-length, Endogenous

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

CPTC-NOTUM-3 Affinity and Kinetics (Bio-Layer Interferometry) 

Result: No Binding

Affinity and binding kinetics of CPTC-NOTUM-3 and human Notum recombinant protein were measured using biolayer interferometry. Recombinant protein was immobilized onto AR2G biosensor using amine coupling. CPTC-NOTUM-3 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without peptide were used as references for background subtraction. Binding data was double referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-NOTUM-3 and human Notum recombinant protein were measured using surface plasmon resonance. Recombinant protein was amine coupled onto a Series S Protein CM5 biosensor chip. CPTC-NOTUM-3 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model. Click image to enlarge

CPTC-NOTUM-3 Affinity and Kinetics (Surface Plasmon Resonance)

Result: High Binding

Affinity and binding kinetics of CPTC-NOTUM-3 and human Notum recombinant protein were measured using surface plasmon resonance. Recombinant protein was amine coupled onto a Series S Protein CM5 biosensor chip. CPTC-NOTUM-3 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  IHC Tissue

CPTC-NOTUM-3 IHC Tissue

Result: Negative

This antibody is not suitable for use in an Immunohistochemistry format as described in SOP M-106.


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-3 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antibody. The antibody recognized the target NOTUM. Click image to enlarge

CPTC-NOTUM-3 automated WB (over-expressed lysate)

Result: Positive

Automated Western Blot using antibody CPTC-NOTUM-3 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antibody. The antibody recognized the target NOTUM.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-3 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody presumably recognized the target NOTUM in the tested cell line lysates. Click image to enlarge

CPTC-NOTUM-3 automated WB (lysate)

Result: Presumed Positive (with additional bands)

Automated Western Blot using antibody CPTC-NOTUM-3 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody presumably recognized the target NOTUM in the tested cell line lysates.


Characterization SOP Files

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Antibody: CPTC-NOTUM-4
Background

Catalog Number:

CPTC-NOTUM-4

Target Antigen:

Notum, Palmitoleoyl-Protein Carboxylesterase

Isotype:

IgG1

Species:

Mouse Monoclonal Antibody

Last Updated:

08/16/2023

Antigen Recognition(s):

Recombinant Full-length, Endogenous

Characterization Data [Compare Characterization Data]
  Affinity Measurement by SPR

CPTC-NOTUM-4 Affinity and Kinetics (Surface Plasmon Resonance)

Result: No Binding

Affinity and binding kinetics of CPTC-NOTUM-4 and human Notum recombinant protein was measured using surface plasmon resonance. Recombinant protein was amine coupled onto a Series S Protein CM5 biosensor chip. CPTC-NOTUM-4 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM was used as analyte. Binding data were double referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  IHC Tissue

CPTC-NOTUM-4 IHC Tissue

Result: Negative

This antibody is not suitable for use in an Immunohistochemistry format as described in SOP M-106.


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-4 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antobody. The antibody recognized the target NOTUM. Click image to enlarge

CPTC-NOTUM-4 automated WB (over-expressed lysate)

Result: Positive

Automated Western Blot using antibody CPTC-NOTUM-4 as primary antibody. The antibody was tested against overexpressed lysate of NOTUM. The same overexpressed lysate was also tested with an anti-Myc antobody. The antibody recognized the target NOTUM.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated Western Blot using antibody CPTC-NOTUM-4 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody presumably recognized the target NOTUM in the tested cell line lysates. Click image to enlarge

CPTC-NOTUM-4 automated WB (lysate)

Result: Presumed Positive (with additional bands)

Automated Western Blot using antibody CPTC-NOTUM-4 as primary antibody. The antibody was tested against cell lysates of SW620 and SF-539. The same lysates was also tested with an anti-CytC antibody. The antibody presumably recognized the target NOTUM in the tested cell line lysates.


Characterization SOP Files

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Antigen: Notum, Palmitoleoyl-Protein Carboxylesterase
Background

NCI Identification Number:

00531

Antigen Name:

Notum, Palmitoleoyl-Protein Carboxylesterase

CPTC Name:

CPTC-NOTUM

Aliases:

Notum, Palmitoleoyl-Protein Carboxylesterase; [Wnt Protein] O-Palmitoleoyl-L-Serine Hydrolase; Palmitoleoyl-Protein Carboxylesterase NOTUM; HNOTUM; Notum Pectinacetylesterase Homolog (Drosophila); Notum Pectinacetylesterase Homolog; Protein Notum Homolog; EC 3.1.1.98

Function:

Enables palmitoleyl hydrolase activity. Involved in negative regulation of Wnt signaling pathway and protein depalmitoleylation. Acts upstream of or within bone development and regulation of bone mineralization. Predicted to be located in endoplasmic reticulum lumen and extracellular region.NOTUM (Notum, Palmitoleoyl-Protein Carboxylesterase) is a Protein Coding gene. Diseases associated with NOTUM include Suppurative Periapical Periodontitis and Hepatocellular Carcinoma. Among its related pathways are Metabolism of proteins and ncRNAs involved in Wnt signaling in hepatocellular carcinoma. Gene Ontology (GO) annotations related to this gene include hydrolase activity and palmitoleyl hydrolase activity.Carboxylesterase that acts as a key negative regulator of the Wnt signaling pathway by specifically mediating depalmitoleoylation of WNT proteins. Serine palmitoleoylation of WNT proteins is required for efficient binding to frizzled receptors (PubMed:25731175).

Chromosomal Localization:

17q25.3

Expression System:

Mammalian

Accession Number:

NP_848588.3

UniProt Accession Number:

Q6P988

DNA Source:

N/A

Immunogen:

Recombinant Domain

Vector Name:

Stably transfected HEK293 cells

Extinction Coefficient:

Buffers:

PBS

Expressed Sequence:

RKTWRRRGQQPPPPPRTEAAPAAGQPVESFPLDFTAVEGNMDSFMAQVKS
LAQSLYPCSAQQLNEDLRLHLLLNTSVTCNDGSPAGYYLKESRGSRRWLL
FLEGGWYCFNRENCDSRYDTMRRLMSSRDWPRTRTGTGILSSQPEENPYW
WNANMVFIPYCSSDVWSGASSKSEKNEYAFMGALIIQEVVRELLGRGLSG
AKVLLLAGSSAGGTGVLLNVDRVAEQLEKLGYPAIQVRGLADSGWFLDNK
QYRHTDCVDTITCAPTEAIRRGIRYWNGVVPERCRRQFQEGEEWNCFFGY
KVYPTLRCPVFVVQWLFDEAQLTVDNVHLTGQPVQEGLRLYIQNLGRELR
HTLKDVPASFAPACLSHEIIIRSHWTDVQVKGTSLPRALHCWDRSLHDSH
KASKTPLKGCPVHLVDSCPWPHCNPSCPTVRDQFTGQEMNVAQFLMHMGF
DMQTVAQPQGLEPSELLGMLSNGS

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

53000

Last Updated:

06/30/2022

Links

Characterization Data

SOPs

No SOPs available.

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