Anti-SMARCA2 / BRM 抗体 [EPR23103-44] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] - BSA and Azide free (AB269872)

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma tissue labeling SMARCA2/BRM with ab240648 at 1/2000 dilution (0.23 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human renal cell carcinoma. The section was incubated with ab240648 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab240648).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] - BSA and Azide free (AB269872)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling SMARCA2/BRM with ab240648 at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear and weak cytoplasmic staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary 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 (ab240648).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SMARCA2 / BRM antibody [EPR23103-44] - BSA and Azide free (AB269872)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling SMARCA2/BRM with ab240648 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A 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 (ab240648).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] - BSA and Azide free (AB269872)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling SMARCA2/BRM with ab240648 at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear and weak cytoplasmic staining in Neuro-2a cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary 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 (ab240648).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] - BSA and Azide free (AB269872)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling SMARCA2/BRM with ab240648 at 1/2000 dilution (0.23 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse cerebrum. The section was incubated with ab240648 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab240648).

• WB

Lab

Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] - BSA and Azide free (AB269872)

This data was developed using the same antibody clone in a different buffer formulation (ab240648).

Lanes 1 - 4 : Merged signal (red and green). Green - ab240648 observed at 200 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab240648 was shown to react with SMARCA2 / BRM in wild-type HeLa cells in Western blot with loss of signal observed in SMARCA2 knockout cell line ab265416 (SMARCA2 knockout cell lysate ab257687). Wild-type HeLa and SMARCA2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab240648 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] (<a href='/products/primary-antibodies/smarca2-brm-antibody-epr23103-44-ab240648'>ab240648</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SMARCA2 knockout HeLa cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 181 kDa

Observed band size: 200 kDa

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