Revision 1

#48585

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Brg1 (D1Q7F) Rabbit mAb4936020 µl220 kDaRabbit IgG
BRD9 (E9R2I) Rabbit mAb5890620 µl80 kDaRabbit IgG
GLTSCR1 (E6I3A) Rabbit mAb4544120 µl205 kDaRabbit IgG
GLTSCR1L (E9K6F) Rabbit mAb9280520 µl140 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Non-Canonical BAF Complex Antibody Sampler Kit provides an economical means of detecting non-canonical BAF complex members using antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

ATP-dependent chromatin remodeling complexes play an essential role in the regulation of various nuclear processes, such as gene expression, DNA replication, and repair (1,2). The SWI/SNF chromatin remodeling complex consists of more than 10 subunits with a single molecule of the ATPase catalytic subunit BRM or BRG1, but not both. The activities of these two subunits drive the disruption of histone-DNA contacts that lead to changes in accessibility of crucial regulatory elements within chromatin (2-5). The BRM/BRG1 containing SWI/SNF complexes are recruited to target promoters by transcription factors, such as nuclear receptors, p53, RB, and BRCA1, to regulate gene activation, cell growth, the cell cycle, and differentiation processes (1,6-9).
GLTSCR1 and its paralog GLTSCR1L, along with BRD9, have been identified as unique subunits of the non-canonical BAF (ncBAF) or GBAF complex. This complex contains either GLTSCR1 or GLTSCR1L instead of an ARID subunit, while also lacking the BAF45, BAF47, and BAF57 subunits. GBAF maps to regions distinct from the PBAF and canonical BAF complexes and has also been shown to be a synthetic lethal target for BAF driven cancers such as synovial sarcomas and rhabdoid tumors (10-12).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using GLTSCR1 (E6I3A) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). GLTSCR1/GLTSCR1L double knockout (KO) cell lysates were isolated and provided by the Kadoch laboratory.
undefined Image 1: Non-Canonical BAF Complex Antibody Sampler Kit
CUT&RUN was performed with NCCIT cells and GLTSCR1 (E6I3A) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across TCF21 gene.
undefined Image 2: Non-Canonical BAF Complex Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using Brg1 (D1Q7F) Rabbit mAb.
undefined Image 3: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
CUT&RUN was performed with NCCIT cells and Brg1 (D1Q7F) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across TNN gene.
undefined Image 4: Non-Canonical BAF Complex Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using BRD9 (E9R2I) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, OVCAR-4 cells are low for BRD9 expression and no BRD9 is detected in the 293T knockout (KO) cells. BRD9 KO cell lysates were isolated and provided by the Kadoch laboratory.
undefined Image 5: Non-Canonical BAF Complex Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 6: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from Jurkat cells, either wild-type (WT) or GLTSCR1/GLTSCR1L double knockout (KO), using GLTSCR1L (E9K6F) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). GLTSCR1/GLTSCR1L double KO cell lysates were isolated and provided by the Kadoch laboratory.
undefined Image 7: Non-Canonical BAF Complex Antibody Sampler Kit
Immunoprecipitation of GLTSCR1 protein from Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is GLTSCR1 (E6I3A) Rabbit mAb. Western blot analysis was performed using GLTSCR1 (E6I3A) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
undefined Image 8: Non-Canonical BAF Complex Antibody Sampler Kit
CUT&RUN was performed with NCCIT cells and GLTSCR1 (E6I3A) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across TCF21 gene (upper) and GATA6 gene (lower).
undefined Image 9: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using Brg1 (D1Q7F) Rabbit mAb (upper) or BRM (D9E8B) XP® Rabbit mAb (lower).
undefined Image 10: Non-Canonical BAF Complex Antibody Sampler Kit
CUT&RUN was performed with NCCIT cells and Brg1 (D1Q7F) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including TNN gene (lower).
undefined Image 11: Non-Canonical BAF Complex Antibody Sampler Kit
Immunoprecipitation of BRD9 protein from HDLM-2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is BRD9 (E9R2I) Rabbit mAb. Western blot analysis was performed using BRD9 (E9R2I) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
undefined Image 12: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from untreated NCCIT cells and either GLTSCR1 (E6I3A) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005.  The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Oct-4 Promoter Primers #4641, SimpleChIP® Human Sox2 Promoter Primers #4649, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
undefined Image 13: Non-Canonical BAF Complex Antibody Sampler Kit
CUT&RUN was performed with NCCIT cells and either GLTSCR1 (E6I3A) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human FOXP2 promoter primers, human YARS2 promoter primers, and human PXDNL promoter 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.
undefined Image 14: Non-Canonical BAF Complex Antibody Sampler Kit
Immunoprecipitation of Brg1 from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Brg1 (D1Q7F) Rabbit mAb. Western blot analysis was performed using Brg1 (D1Q7F) Rabbit mAb.
undefined Image 15: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
CUT&RUN was performed with NCCIT cells and either Brg1 (D1Q7F) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA2 Promoter Primers #5517, and human ITM2A upstream 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.
undefined Image 16: Non-Canonical BAF Complex Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d followed by treatment with β-estradiol (10 nM, 45 min) and Brg1 (D1Q7F) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across the ENSA gene.
undefined Image 17: Non-Canonical BAF Complex Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d followed by treatment with β-estradiol (10 nM, 45 min) and Brg1 (D1Q7F) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across ESR1 (upper), a known target gene of Brg1 (see additional figure containing ChIP-qPCR data), and ENSA gene (lower).
undefined Image 18: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d followed by treatment with β-estradiol (10 nM, 45 min) and either Brg1 (D1Q7F) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
undefined Image 19: Non-Canonical BAF Complex Antibody Sampler Kit
Simple Western™ analysis of lysates (1.0 mg/mL) from HDLM-2 cells using BRD9 (E9R2I) Rabbit mAb #58906. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 20: Non-Canonical BAF Complex Antibody Sampler Kit
Immunoprecipitation of GLTSCR1L protein from Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is GLTSCR1L (E9K6F) Rabbit mAb. Western blot analysis was performed using GLTSCR1L (E9K6F) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
undefined Image 21: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using GLTSCR1L (E9K6F) Rabbit mAb.
undefined Image 22: Non-Canonical BAF Complex Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.