BAF170 Recombinant Rabbit Monoclonal Antibody [PSH12-00]
cat.: HA723413
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, IP, ChIP
Clonality: Monoclonal
Clone number: PSH12-00
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 133 kDa
Isotype: IgG
Immunogen: Recombinant protein within human BAF170 aa 915-1,214.
Positive control: K-562 cell lysate, A431 cell lysate, MCF7 cell lysate, HeLa cell lysate, SW620 cell lysate, SW680 cell lysate, PANC cell lysate, C2C12 cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, human brain tissue, human stomach tissue, mouse brain tissue, mouse stomach tissue, rat brain tissue, rat stomach tissue, HeLa, Neuro-2a.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  IP
  ChIP
1:5,000
1:1,000
1:500
1-2μg/sample
Use 5 μg for 25 μg of chromatin.
Uniprot #: SwissProt: Q8TAQ2 Human | Q6PDG5 Mouse
Entrez Gene: 362815 Rat
Alternative names: BAF 170 BAF170 BRG1 associated factor 170 BRG1-associated factor 170 Chromatin remodeling complex BAF170 subunit CRACC 2 CRACC2 Mammalian chromatin remodeling complex BRG1 associated factor 170 Rsc 8 Rsc8 SMARCC 2 Smarcc2 SMRC2_HUMAN SWI/SNF complex 170 kDa subunit SWI/SNF complex subunit SMARCC2 SWI/SNF related matrix associated actin dependent regulator of chromatin c2 SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily c member 2 SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily C member 2 SWI3 like protein
Images
HA723413_1.jpg Fig1: Western blot analysis of BAF170 on different lysates with Rabbit anti-BAF170 antibody (HA723413) at 1/5,000 dilution.

Lane 1: K-562 cell lysate (20 µg/Lane)
Lane 2: A431 cell lysate (20 µg/Lane)
Lane 3: MCF7 cell lysate (20 µg/Lane)
Lane 4: HeLa cell lysate (20 µg/Lane)
Lane 5: SW620 cell lysate (20 µg/Lane)
Lane 6: SW680 cell lysate (20 µg/Lane)
Lane 7: PANC cell lysate (20 µg/Lane)
Lane 8: C2C12 cell lysate (20 µg/Lane)
Lane 9: NIH/3T3 cell lysate (20 µg/Lane)
Lane 10: Neuro-2a cell lysate (20 µg/Lane)
Lane 11: PC-12 cell lysate (20 µg/Lane)
Lane 12: Mouse brain tissue lysate (40 µg/Lane)
Lane 13: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 133 kDa
Observed band size: 162 kDa

Exposure time: 25 seconds; 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 (HA1001) at 1/1,000 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.
HA723413_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-BAF170 antibody (HA723413) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723413) 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.
HA723413_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-BAF170 antibody (HA723413) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723413) 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.
HA723413_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-BAF170 antibody (HA723413) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723413) 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.
HA723413_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-BAF170 antibody (HA723413) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723413) 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.
HA723413_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-BAF170 antibody (HA723413) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723413) 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.
HA723413_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-BAF170 antibody (HA723413) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723413) 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.
HA723413_8.jpg Fig8: Immunocytochemistry analysis of HeLa cells labeling BAF170 with Rabbit anti-BAF170 antibody (HA723413) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-BAF170 antibody (HA723413) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1126) 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 (HA601187, 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.
HA723413_9.jpg Fig9: Immunocytochemistry analysis of Neuro-2a cells labeling BAF170 with Rabbit anti-BAF170 antibody (HA723413) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-BAF170 antibody (HA723413) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1126) 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 (HA601187, 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.
HA723413_10.jpg Fig10: BAF170 was immunoprecipitated from 0.2 mg MCF7 cell lysate with HA723413 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723413 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: MCF7 cell lysate (input)
Lane 2: HA723413 IP in MCF7 cell lysate
Lane 3: Rabbit IgG instead of HA723413 in MCF7 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 30 seconds; ECL: K1801
HA723413_11.jpg Fig11: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with BAF170 (HA723413) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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