ET1611-85

BRG1 Recombinant Rabbit Monoclonal Antibody [SN20-03]

cat.: ET1611-85

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC, IHC-Fr
Clonality:	Monoclonal
Clone number:	SN20-03

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 185 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human BRG1 aa 240-280.
Positive control:	HeLa, HeLa cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, HepG2, NIH/3T3, human tonsil tissue, human kidney tissue, human breast carcinoma tissue, mouse testis tissue, mouse colon tissue, mouse kidney tissue, mouse epididymis tissue, rat colon tissue, rat kidney tissue, rat brain tissue, mouse hippocampus tissue, mouse cerebral cortex tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P IP FC IHC-Fr	1:20,000-1:50,000 1:200-1:1,000 1:50-1:200 1:500-1:1,000 Use at an assay dependent concentration. 1:1,000 1:100
Uniprot #:	SwissProt: P51532 Human \| Q3TKT4 Mouse \| Q8K1P7 Rat
Alternative names:	ATP dependent helicase SMARCA4 ATP-dependent helicase SMARCA4 BAF 190 BAF190 BAF190A Brahma protein like 1 BRG1 BRG1 associated factor 190A BRG1 protein BRG1-associated factor 190A BRM/SWI2 related gene 1 Global transcription activator homologous sequence global transcription activator snf2l4 Homeotic gene regulator hSNF2b Mitotic growth and transcription activator MRD16 Nuclear protein GRB1 Protein brahma homolog 1 Protein BRG-1 Protein BRG1 RTPS2 SMARC A4 SMARCA4 SMCA4_HUMAN SNF2 SNF2 beta SNF2 like 4 SNF2-beta SNF2B SNF2L4 SNF2LB Sucrose nonfermenting like 4 SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily A member 4 SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4 SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4 SWI2 Transcription activator BRG1

Images

Fig1: Immunocytochemistry analysis of HeLa cells labeling BRG1 with Rabbit anti-BRG1 antibody (ET1611-85) at 1/1,000 dilution and competitor's antibody at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BRG1 antibody (ET1611-85) at 1/1,000 dilution and competitor's antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Fig2: Western blot analysis of BRG1 on different lysates with Rabbit anti-BRG1 antibody (ET1611-85) at 1/50,000 dilution and competitor's antibody at 1/10,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: K-562 cell lysate
Lane 3: A549 cell lysate (negative)
Lane 4: NIH/3T3 cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: C6 cell lysate
Lane 7: PC-12 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 185 kDa
Observed band size: 200 kDa

Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-85) at 1/50,000 dilution and competitor's antibody at 1/10,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of BRG1 on different lysates with Rabbit anti-BRG1 antibody (ET1611-85) at 1/2,000 dilution.

Lane 1: HeLa-si NT cell lysate
Lane 2: HeLa-si BRG1 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 185 kDa
Observed band size: 200 kDa

Exposure time: 20 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-85) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig4: Immunocytochemistry analysis of HepG2 cells labeling BRG1 with Rabbit anti-BRG1 antibody (ET1611-85) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BRG1 antibody (ET1611-85) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling BRG1 with Rabbit anti-BRG1 antibody (ET1611-85) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BRG1 antibody (ET1611-85) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig8: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig9: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig10: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig11: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig12: Immunohistochemical analysis of paraffin-embedded mouse epididymis tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig13: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig14: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-BRG1 antibody (ET1611-85) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-85) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig15: Immunofluorescence analysis of paraffin-embedded rat brain tissue labeling BRG1 with Rabbit anti-BRG1 antibody (ET1611-85) at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1611-85, green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig16: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling BRG1 with Rabbit anti-BRG1 antibody (ET1611-85). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1611-85, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
	Fig17: Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling BRG1 with Rabbit anti-BRG1 antibody (ET1611-85). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1611-85, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

Fig18: Flow cytometric analysis of HeLa cells labeling BRG1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-85, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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