Anti-Proteasome 20S LMP2 抗体 [EPR22042]

• Flow Cyt (Intra)

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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.

• IHC-P

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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).

• IHC-P

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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).

• IHC-P

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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).

• ICC/IF

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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.

• IP

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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.

All lanes:

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

Predicted band size: 23 kDa

Observed band size: 21 kDa,23 kDa

false

• IHC-P

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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).

• Flow Cyt (Intra)

Supplier Data

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.

• IHC-P

Supplier Data

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).

• ICC/IF

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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.

• WB

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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 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).

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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 21 kDa,23 kDa

false

Exposure time: 59s

• WB

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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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 21 kDa,23 kDa

false

• WB

CiteAb

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)

false