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

Rabbit Recombinant Monoclonal Proteasome 20S LMP2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

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Images

Immunoprecipitation - Anti-Proteasome 20S LMP2 antibody [EPR22042] (AB242061), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Tested	Tested	Tested
Rat	Expected	Tested	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -
Species Human	Dilution info 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information PSMB9

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 (PubMed:33727065, PubMed:34819510). 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.

Alternative names

LMP2, PSMB6i, RING12, PSMB9, Proteasome subunit beta type-9, Low molecular mass protein 2, Macropain chain 7, Multicatalytic endopeptidase complex chain 7, Proteasome chain 7, Proteasome subunit beta-1i, Really interesting new gene 12 protein

Recommended products

Rabbit Recombinant Monoclonal Proteasome 20S LMP2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR22042
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The proteasome LMP2 also known as proteasome subunit beta type-1 or PSMB9 is a component important in the functioning of the 20S proteasome complex. This molecular entity with a mass of approximately 23 kDa is predominantly expressed in immune cells including lymphocytes. LMP2 incorporates into the immunoproteasome a specialized variant of the proteasome that participates prominently in the processing of class I MHC peptides enhancing the immune response.

Biological function summary

LMP2 is integral to protein catabolism and regulation of the cellular protein homeostasis. As part of the larger proteasome complex the LMP2 protein functions in degradative pathways that recycle amino acids from ubiquitin-tagged proteins. This proteasome takes part in antigen processing helping convert proteins into peptides that are presented on the surface of cells for immune system recognition.

Pathways

LMP2 is significantly involved in the ubiquitin-proteasome pathway and immune system processes. It interacts with other immunoproteasome subunits like LMP7 and MECL-1 which work together to enhance the proteolytic capacity for antigen presentation. The presence of LMP2 in these pathways demonstrates its role in adaptive immunity contributing to the clearance of misfolded or damaged proteins and therefore maintaining cell health.

Associated diseases and disorders

LMP2 associates with immune-related conditions and some cancer types. Dysregulation of LMP2 expression or function can impact antigen presentation potentially leading to autoimmune disorders due to inappropriate immune activation. Moreover alterations in LMP2 expression have been observed in certain cancers such as melanoma where it affects proteasome activity and influences tumor progression. In these contexts interactions with proteins like TAP1 and TAP2 which aid in peptide transport mark its relevance in disease pathogenesis.

Product promise

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13 product images

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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab242061 in THP-1 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 10 seconds.
All lanes: Immunoprecipitation - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)
Predicted band size: 23 kDa
Observed band size: 21 kDa, 23 kDa
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (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) (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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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)
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).
All lanes: Western blot - 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: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 21 kDa, 23 kDa
Western blot - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)
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 Anti-GAPDH antibody [EPR16891] - Loading Control 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 (Goat Anti-Rabbit IgG H&L (HRP) 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).
All lanes: Western blot - 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: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 21 kDa, 23 kDa
Exposure time: 59s
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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-Proteasome 20S LMP2 antibody [EPR22042] (ab242061)
Proteasome 20S LMP2 western blot using anti-Proteasome 20S LMP2 antibody [EPR22042] ab242061. Publication image and figure legend from Ye, J., Yin, Y., et al., 2020, Aging Cell, PubMed 31668016.

ab242061 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab242061 please see the product overview.
Overexpressing hTau disrupts PA28γ‐20S proteasome complex formation and downregulating PKAR2α or upregulating PA28γ attenuates hTau‐induced PKA/CREB inhibition. (a) Overexpressing hTau increases association of PKAR2α with PA28γ. The cultured hippocampal neurons infected with lenti‐syn‐hTau‐mCherry virus for 7 days, and PA28γ was co‐immunoprecipitated by anti‐PA28γ antibody and detected by Western blot using anti‐PKAR2α and anti‐PA28γ antibody. For all co‐immunoprecipitations, results are received from three independent experiments. (b) Overexpressing hTau decreases association of PKAR2α with 20S proteasome subunit. PKAR2α was co‐immunoprecipitated using anti‐PKAR2α antibody and detected by Western blot using anti‐20S proteasome subunit and anti‐PKAR2α antibody. (c) Overexpressing hTau decreases association of PA28γ with 20S proteasome subunit. 20S proteasome subunit was co‐immunoprecipitated using anti‐20S protease subunit and detected by Western blot using anti‐PA28γ antibody and anti‐20S proteasome subunit. (d) Tau dose not bind to PA28γ. The cultured hippocampal neuron extract was co‐immunoprecipitated using anti‐PA28γ, and tau was not detected in the precipitates by Tau5. (e, f) Downregulating PKAR2α by shRNA attenuates hTau‐induced CREB dephosphorylation. The primary cultured hippocampal neurons (5 div) were co‐infected with hTau and shPKAR2α lentivirus. After 7 days, the indicated proteins were measured by Western blot. (g, h) Upregulating PA28γ attenuates hTau‐induced PKAR2α elevation and CREB dephosphorylation. The hTau and PA28γ lentivirus were co‐infected in primary cultured hippocampal neurons (5 div) for 7 days, and the indicated proteins were measured by Western blot. Data were expressed as mean ± SEM. For Western blot, the n number was 9 (from nine different batches of cultured hippocampal neurons). *p < .05, **p < .01, versus Vec, &p < .05, versus hTau in (f), #p < .05, ##p < .01, versus hTau in (h)