Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• IP

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Immunoprecipitation - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

All lanes:

Immunoprecipitation - Anti-Proteasome 20S LMP2 antibody [EPR22042] (<a href='/en-us/products/primary-antibodies/proteasome-20s-lmp2-antibody-epr22042-ab242061'>ab242061</a>)

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] - BSA and Azide free (AB243556)

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).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

• WB

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Western blot - Anti-Proteasome 20S LMP2 antibody [EPR22042] - BSA and Azide free (AB243556)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab242061).

All lanes:

Western blot - Anti-Proteasome 20S LMP2 antibody [EPR22042] (<a href='/en-us/products/primary-antibodies/proteasome-20s-lmp2-antibody-epr22042-ab242061'>ab242061</a>) 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='/en-us/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