Recombinant Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22042] to Proteasome 20S LMP2
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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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, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Human Proteasome 20S LMP2 aa 1 to the C-terminus. The exact sequence is proprietary.
Database link: P28065 -
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: Ramos and RAW 264.7 cells. IP: THP-1 whole cell lysate.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22042 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
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Recombinant Protein
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Related Products
Applications
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 |
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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. | |
Flow Cyt | 1/500. | |
IP | 1/30. |
Target
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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
modificationsAutocleaved. The resulting N-terminal Thr residue of the mature subunit is responsible for the nucleophile proteolytic activity. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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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
see all -
Alternative names
- Beta1i antibody
- Large multifunctional peptidase 2 antibody
- Large multifunctional protease 2 antibody
see all
Images
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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 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.
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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.
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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. -
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. -
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).
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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 secondsab242061 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).
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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).
Protocols
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.
Datasheets and documents
Certificate of Compliance
References (1)
ab242061 has been referenced in 1 publication.
- 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