Recombinant Anti-Proteasome 20S LMP2 antibody [EPR22042] KO Tested (ab242061)

Product name

Anti-Proteasome 20S LMP2 antibody [EPR22042]
See all Proteasome 20S LMP2 primary antibodies
Description

Rabbit monoclonal [EPR22042] to Proteasome 20S LMP2
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Wild-type HAP1. Ramos, C6, NIH/3T3, HL-60, Raji and RAW 264.7 whole cell lysate. IHC-P: Human colon cancer, cerebrum and prostatic hyperplasia tissue. Mouse and rat liver tissue. ICC/IF: Ramos and RAW 264.7 cells. Flow Cyt (intra): Ramos and RAW 264.7 cells. IP: THP-1 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22042
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
- Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (ab243556)
Compatible Secondaries
Immunohistochemistry reagents
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Proteasome 20S LMP2 protein (ab105592)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab242061 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Predicted molecular weight: 23 kDa.
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/100.
IP		1/30.
Flow Cyt (Intra)		1/500.

Notes
WB 1/1000. Predicted molecular weight: 23 kDa.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/30.
Flow Cyt (Intra) 1/500.

Function

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. This subunit is involved in antigen processing to generate class I binding peptides. Replacement of PSMB6 by PSMB9 increases the capacity of the immunoproteasome to cleave model peptides after hydrophobic and basic residues.
Sequence similarities

Belongs to the peptidase T1B family.
Developmental stage

Highly expressed in immature dendritic cells (at protein level).
Post-translational
modifications

Autocleaved. The resulting N-terminal Thr residue of the mature subunit is responsible for the nucleophile proteolytic activity.
Cellular localization

Cytoplasm. Nucleus.
Target information above from: UniProt accession P28065 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 5698 Human
- Entrez Gene: 16912 Mouse
- Entrez Gene: 24967 Rat
- Omim: 177045 Human
- SwissProt: P28065 Human
- SwissProt: P28076 Mouse
- SwissProt: P28077 Rat
- Unigene: 654585 Human
- Unigene: 390983 Mouse
- Unigene: 13686 Rat
see all
Alternative names
- Beta1i antibody
- Large multifunctional peptidase 2 antibody
- Large multifunctional protease 2 antibody
- LMP 2 antibody
- LMP2 antibody
- Low molecular mass protein 2 antibody
- Macropain chain 7 antibody
- MGC70470 antibody
- Multicatalytic endopeptidase complex chain 7 antibody
- OTTHUMP00000062982 antibody
- Proteasome (prosome macropain) subunit beta type 9 antibody
- proteasome (prosome, macropain) subunit, beta type, 9 (large multifunctional peptidase 2) antibody
- Proteasome beta 9 subunit antibody
- Proteasome catalytic subunit 1i antibody
- Proteasome chain 7 antibody
- Proteasome related gene 2 antibody
- Proteasome subunit beta 6i antibody
- Proteasome subunit beta type 9 antibody
- Proteasome subunit beta type-9 antibody
- Proteasome subunit beta-1i antibody
- PSB9_HUMAN antibody
- PSMB 9 antibody
- PSMB6i antibody
- PSMB9 antibody
- Really interesting new gene 12 protein antibody
- RING 12 antibody
- RING12 antibody
- RING12 protein antibody
see all

Immunoprecipitation - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Proteasome 20S LMP2 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate with ab242061 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab242061 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: THP-1 whole cell lysate 10 µg (Input).
Lane 2: ab242061 IP in THP-1 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab242061 in THP-1 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 10 seconds.
Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cell line labeling Proteasome 20S LMP2 with ab242061 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Ramos (Human Burkitt's lymphoma B lymphocyte) cell line labeling Proteasome 20S LMP2 with ab242061 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling Proteasome 20S LMP2 with ab242061 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in RAW 264.7 cells. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.
Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling Proteasome 20S LMP2 with ab242061 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in Ramos cells. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.
Western blot - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

All lanes : Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate at 10 µg
Lane 2 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg
Lane 3 : HL-60 (human Acute Promyelocytic Leukemia promyeloblast), whole cell lysate at 20 µg
Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 23 kDa
Observed band size: 21,23 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-2: 15 seconds, Lanes 3:3 minutes, Lane 4: 59 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID:22355695; PMID:15284441).
Western blot - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

All lanes : Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061) at 1/1000 dilution

Lane 1 : Wild-type HAP1 at 40 µg
Lane 2 : Proteasome 20S LMP2 knockout HAP1 whole cell lysate at 40 µg
Lane 3 : Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg
Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 23 kDa
Observed band size: 21,23 kDa why is the actual band size different from the predicted?

Exposure time: 59 seconds

ab242061 was shown to specifically react with Proteasome 20S LMP2 in wild-type HAP1 cells as signal was lost in Proteasome 20S LMP2 knockout cells. Wild-type and Proteasome 20S LMP2 knockout samples were subjected to SDS-PAGE. ab242061 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument using the ECL technique.

The expression profile observed is consistent with what has been described in the literature (PMID:22355695; PMID:15284441).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Proteasome 20S LMP2 with ab242061 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in rat liver (PMID:8365398) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Proteasome 20S LMP2 with ab242061 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in mouse liver (PMID:8365398) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Proteasome 20S LMP2 with ab242061 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in human cerebrum (PMID:20174631) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Proteasome 20S LMP2 with ab242061 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in human colon cancer (PMID:24040045) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

Immunohistochemical analysis of paraffin-embedded human prostatic hyperplasia tissue labeling Proteasome 20S LMP2 with ab242061 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in human prostatic hyperplasia (PMID:22677907) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab242061? Please let us know so that we can cite the reference in this datasheet.

ab242061 has been referenced in 2 publications.

Ye J et al. Tau inhibits PKA by nuclear proteasome-dependent PKAR2a elevation with suppressed CREB/GluA1 phosphorylation. Aging Cell 19:e13055 (2020). PubMed: 31668016
Du SH et al. Co-Inhibition of the Immunoproteasome Subunits LMP2 and LMP7 Ameliorates Immune Thrombocytopenia. Front Immunol 11:603278 (2020). PubMed: 33552061

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Immunohistochemistry reagents

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Developmental stage

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As

Post-translational
modifications