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Proteasome Subunit Beta 11 Peptide 1

Antibody: CPTC-PSMB11 (mouse)-1
Background

Catalog Number:

CPTC-PSMB11 (mouse)-1

RRID:

AB_2943353

Target Antigen:

Proteasome Subunit Beta 11 Peptide 1

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

07/03/2024

Antigen Recognition(s):

Peptide

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

CPTC-PSMB11-1 Affinity and Kinetics (Bio-Layer Interferometry)

Result: Negative

Affinity and binding kinetics of CPTC-PSMB11-1 and BSA-conjugated peptide, "VEQEEVTPEDCAIIMKTETM" were measured using biolayer interferometry. BSA-conjugated peptide was amine coupled on AR2G- biosensor. CPTC-PSMB11-1 rabbit antibody was used as analyte and was titrated at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM. Buffer only and biosensors immobilized without peptide were used as references for background subtraction. All binding data were double referenced and analyzed globally using a 1:2 bivalent fitting model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-PSMB11-1 antibody and BSA-conjugated peptide "VEQEEVTPEDCAIIMKTETM" were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-PSMB11-1 rabbit antibody was used as analyte and was titrated at 256 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM. All binding data were double-referenced and analyzed globally using a 1:2 bivalent fitting model.  Click image to enlarge

CPTC-PSMB11-1 Affinity and Kinetics (Surface Plasmon Resonance)

Result: Positive

Affinity and binding kinetics of CPTC-PSMB11-1 antibody and BSA-conjugated peptide "VEQEEVTPEDCAIIMKTETM" were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-PSMB11-1 rabbit antibody was used as analyte and was titrated at 256 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM. All binding data were double-referenced and analyzed globally using a 1:2 bivalent fitting model. 


Characterization SOP Files

  Flow Cytometry

CPTC-PSMB11-1 Flow Cytometry

Result: Negative

Flow cytometric analysis of PSMB11 expression using CPTC-PSMB11-1 rabbit antibody. Mouse EL4.IL-2 cells were fixed and then stained with CPTC-PSMB11-1 (solid blue) or concentration-matched rabbit isotype control (dashed blue). Human Hs 202.Th cells were fixed and then stained with CPTC-PSMB11-1 (solid green) or concentration-matched rabbit isotype control (dashed green). A BV421 conjugated goat anti-rabbit IgG (H+L) was used as a secondary antibody. All data were analyzed using FlowJo. 


Characterization SOP Files

  IP

CPTC-PSMB11-1 Immunoprecipitation (tissue lysates)

Result: Negative

Immunoprecipitation using CPTC-PSMB11-1 as precipitating antibody against human thymus tissue (negative control) and mouse thymus tissue (positive control) lysates . The eluate was screened in SW using CPTC-PSMB11-1 as detection antibody. The antibody is not able to pull down the target protein (mouse PSMB11).


Characterization SOP Files

  Immunofluorescence

CPTC-PSMB11-1 Immunofluorescence

Result: Negative

Immunofluorescence staining using CPTC-PSMB11-1 as primary antibody against mouse EL4.IL-2 and human Hs. 202.Th cells. Both cell lines show no localization of PSMB11 protein.


Characterization SOP Files

  Immunofluorescence

CPTC-PSMB11-1 Immunofluorescence

Result: Negative

Immunofluorescence staining using CPTC-PSMB11-1 as primary antibody against engineered MEF beta 5i (positive) and MEF beta 5i KO (negative) cells. CPTC-PSMB11-1 ab shows no PSMB11 localization in neither cell lines.


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA using CPTC-PSMB11-1 as primary rabbit antibody against BSA-conjugated peptide "VEQEEVTPEDCAIIMKTETM",coated on the plate and detected using goat-anti rabbit antibody and TMB. Click image to enlarge

CPTC-PSMB11-1 Indirect ELISA

Result: Positive

Indirect ELISA using CPTC-PSMB11-1 as primary rabbit antibody against BSA-conjugated peptide "VEQEEVTPEDCAIIMKTETM",coated on the plate and detected using goat-anti rabbit antibody and TMB.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-PSMB11-1 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
Click to enlarge image Simple Western using CPTC-PSMB11-1 as primary antibody against human thymus tissue, mouse thymus tissue, mouse lung tissue, mouse liver tissue, and mouse spleen tissue. The antibody detected the target protein (~33 KDa) only in the mouse thymus tissue, as expected. The same cell lysates were also probed for Cytochrome C, which was detected at different level of expression in each tested cell. Click image to enlarge

CPTC-PSMB11-1 Automated Western (Tissue)

Result: Positive

Simple Western using CPTC-PSMB11-1 as primary antibody against human thymus tissue, mouse thymus tissue, mouse lung tissue, mouse liver tissue, and mouse spleen tissue. The antibody detected the target protein (~33 KDa) only in the mouse thymus tissue, as expected. The same cell lysates were also probed for Cytochrome C, which was detected at different level of expression in each tested cell.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western Blot using CPTC-PSMB11-1 as primary antibody against human thymus tissue, mouse thymus tissue, mouse lung tissue, mouse liver tissue, and mouse spleen tissue. The antibody detected the target protein (~33 KDa) only in the mouse thymus tissue, as expected. The same cell lysates were also probed for Cytochrome C, which was detected at different level of expression in each tested cell. Click image to enlarge

CPTC-PSMB11-1 Western Blot (Tissue Lysates)

Result: Positive

Western Blot using CPTC-PSMB11-1 as primary antibody against human thymus tissue, mouse thymus tissue, mouse lung tissue, mouse liver tissue, and mouse spleen tissue. The antibody detected the target protein (~33 KDa) only in the mouse thymus tissue, as expected. The same cell lysates were also probed for Cytochrome C, which was detected at different level of expression in each tested cell.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western Blot using CPTC-PSMB11-1 as primary antibody against the cell lysates of mouse thymus (EL4.IL2), mouse lymphoma cells (EL4), and human Thymus cells (Hs.202.Th). The antibody does not recognize the target protein in the tested lysates. The same cell lysates were also probed for Cytochrome C, which was detected at different level of expression in each tested cell. Click image to enlarge

CPTC-PSMB11-1 Western Blot (Cell Lysates)

Result: Negative

Western Blot using CPTC-PSMB11-1 as primary antibody against the cell lysates of mouse thymus (EL4.IL2), mouse lymphoma cells (EL4), and human Thymus cells (Hs.202.Th). The antibody does not recognize the target protein in the tested lysates. The same cell lysates were also probed for Cytochrome C, which was detected at different level of expression in each tested cell.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate

CPTC-PSMB11-1 Single Cell Western Blot (Cell Lysate)

Result: Negative

Single cell western blot using CPTC-PSMB11-1 as a primary antibody against mouse EL4.IL-2 and human Hs. 202.Th cell lysates. PSMB11 protein expression was not detected in neither cell lysates


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate

CPTC-PSMB11-1 Single Cell Western Blot (Cell Lysate)

Result: Negative

Single cell western blot using CPTC-PSMB11-1 as a primary antibody against engineered MEF beta 5i (positive) and MEF beta 5i KO (negative) cell lysates. PSMB11 protein expression was not detected in neither cell lysates.


Characterization SOP Files

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Antigen: Proteasome Subunit Beta 11 Peptide 1
Background

NCI Identification Number:

00548

Antigen Name:

Proteasome Subunit Beta 11 Peptide 1

CPTC Name:

CPTC-PSMB11 (mouse) Peptide 1

Aliases:

Proteasome Subunit Beta 11; Proteasome (Prosome, Macropain) Subunit, Beta Type, 11; Proteasome Subunit Beta Type-11; Proteasome Subunit Beta-5t; EC 3.4.25.1; Beta5t; Proteasome Subunit Beta 5T; BETA5T

Function:

Proteasomes generate peptides that are presented by major histocompatibility complex (MHC) I molecules to other cells of the immune system. Proteolysis is conducted by 20S proteasomes, complexes of 28 subunits arranged as a cylinder in 4 heteroheptameric rings: alpha-1 to -7, beta-1 to -7, beta-1 to -7, and alpha-1 to -7. The catalytic subunits are beta-1 (PSMB6; MIM 600307), beta-2 (PSMB7; MIM 604030), and beta-5 (PSMB5; MIM 600306). Three additional subunits, beta-1i (PSMB9; MIM 177045), beta-2i (PSMB10; MIM 176847), and beta-5i (PSMB8; MIM 177046), are induced by gamma-interferon (IFNG; MIM 147570) and are preferentially incorporated into proteasomes to make immunoproteasomes. PSMB11, or beta-5t, is a catalytic subunit expressed exclusively in cortical thymic epithelial cells.PSMB11 (Proteasome Subunit Beta 11) is a Protein Coding gene. Diseases associated with PSMB11 include Encapsulated Thymoma and Thymic Dysplasia. Among its related pathways are Regulation of activated PAK-2p34 by proteasome mediated degradation and RAF/MAP kinase cascade. Gene Ontology (GO) annotations related to this gene include threonine-type endopeptidase activity. An important paralog of this gene is PSMB8.The proteasome is a multicatalytic proteinase complex which is characterized by its ability to cleave peptides with Arg, Phe, Tyr, Leu, and Glu adjacent to the leaving group at neutral or slightly basic pH. The proteasome has an ATP-dependent proteolytic activity. Incorporated instead of PSMB5 or PSMB8, this unit reduces the chymotrypsin-like activity of the proteasome (By similarity). Plays a pivotal role in development of CD8-positive T cells (By similarity).

Chromosomal Localization:

14q11.2

Accession Number:

UniProt Accession Number:

Q8BG41

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

VEQEEVTPEDCAIIMKTETM

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

2200

Last Updated:

10/19/2023

Links

Characterization Data

SOPs

No SOPs available.

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