Anti-SMARCA6 antibody [EPR25295-62]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling SMARCA6 with ab300040 at 1/500 (1.06 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human testis (PMID : 21349825). The section was incubated with ab300040 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 LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized LNCaP (Human prostate carcinoma epithelial cell, Left) / MOLT-4 (human lymphoblastic leukemia T lymphoblast, Right) cells labeling SMARCA6 with ab300040 at 1/500 dilution (0.1 μg) (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, ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression : LNCaP(PMID : 10910076).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling SMARCA6 with ab300040 at 1/50 dilution (1 μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling SMARCA6 with ab300040 at 1/500 dilution (1.06 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on rat testis (PMID : 21349825). The section was incubated with ab300040 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 LeicaDS9800 (BOND™ Polymer Refine Detection).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized NIH/3T3 (mouse embryonic fibroblast) cells labelling SMARCA6 with ab300040 at 1/50 dilution (10.6 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/ml) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 μg/mL) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling SMARCA6 with ab300040 at 1/500 (1.06 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on mouse testis (PMID : 21349825). The section was incubated with ab300040 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 LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

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Immunoprecipitation - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

SMARCA6 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg with ab300040 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300040 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg

Lane 2 : ab300040 in NIH/3T3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300040 in NIH/3T3 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-SMARCA6 antibody [EPR25295-62] (ab300040)

Predicted band size: 97 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

SMARCA6 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma b lymphocyte) whole cell lysate 10 μg with ab300040 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300040 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Raji (human Burkitt's lymphoma b lymphocyte) whole cell lysate 10 μg

Lane 2 : ab300040 in Raji whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300040 in Raji whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-SMARCA6 antibody [EPR25295-62] (ab300040)

Predicted band size: 97 kDa

false

• WB

Unknown

Western blot - Anti-SMARCA6 antibody [EPR25295-62] (AB300040)

Blocking / Diluting buffer and concentration :

5% NFDM/TBST

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 10910076).
Low expression cell line : LNCaP, RPMI-8226(PMID : 10910076).

All lanes:

Western blot - Anti-SMARCA6 antibody [EPR25295-62] (ab300040) at 1/1000 dilution

Lane 1:

MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate at 20 µg

Lane 2:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 4:

RPMI-8226 (human plasmacytoma, myeloma b lymphocyte) whole cell lysate at 20 µg

Lane 5:

Mouse thymus tissue lysate at 20 µg

Lane 6:

Rat thymus tissue lysate at 20 µg

Lane 7:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 8:

RAW 264.7 (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: 97 kDa

Observed band size: 32-100 kDa

false

Exposure time: 3min