Anti-MBD2 antibody [EPR18361] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

Intracellular Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling MBD2 with purified ab188474 at 1/70 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188474).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling MBD2 with ab188474 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HepG2 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse (AlexaFluor^®594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab188474 at 1/250 dilution, followed by Goat Anti-Mouse (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188474).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human colon tissue tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188474).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human gastric cancer tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188474).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

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Immunoprecipitation - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

MBD2 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab188474 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab188474 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1 : HeLa whole cell lysate 10µg (Input).
Lane 2 : ab188474 IP in HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab188474 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188474).

All lanes:

Immunoprecipitation - Anti-MBD2 antibody [EPR18361] (<a href='/en-us/products/primary-antibodies/mbd2-antibody-epr18361-ab188474'>ab188474</a>)

Predicted band size: 43 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat colon tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188474).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse stomach tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188474).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-MBD2 antibody [EPR18361] - BSA and Azide free (AB224274)

This WB data was generated using the same anti-MBD2 antibody clone [EPR18361] in a different buffer formulation (cat# ab188474).

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : MBD2 knockout HAP1 cell lysate (20 μg)
Lane 3 : A375 cell lysate (20 μg)
Lane 4 : Human brain tissue lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab188474 observed at 32 & 49 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab188474 was shown to specifically react with MBD2 when MBD2 knockout samples were used. Wild-type and MBD2 knockout samples were subjected to SDS-PAGE. ab188474 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ( ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-MBD2 antibody [EPR18361] (<a href='/en-us/products/primary-antibodies/mbd2-antibody-epr18361-ab188474'>ab188474</a>)

Predicted band size: 43 kDa

false