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48585
Non-Canonical BAF Complex Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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Non-Canonical BAF Complex Antibody Sampler Kit #48585

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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.
Western blot analysis of extracts from various cell lines using GLTSCR1L (E9K6F) Rabbit mAb.
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.
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.
Western blot analysis of extracts from various cell lines using Brg1 (D1Q7F) Rabbit mAb.
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.
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.
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.
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.
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.
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).
Western blot analysis of extracts from various cell lines using Brg1 (D1Q7F) Rabbit mAb (upper) or BRM (D9E8B) XP® Rabbit mAb (lower).
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).
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.
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.
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.
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.
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.
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.
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).
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.
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To Purchase # 48585
製品# サイズ 数量 価格 在庫
48585T		 1 Kit  (4 x 20 microliters)

Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Brg1 (D1Q7F) Rabbit mAb 49360 20 µl
  • WB
  • IP
  • ChIP
  • C&R
 H M R Mk 220 Rabbit IgG
BRD9 (E9R2I) Rabbit mAb 58906 20 µl
  • WB
  • IP
 H Mk 80 Rabbit IgG
GLTSCR1 (E6I3A) Rabbit mAb 45441 20 µl
  • WB
  • IP
  • ChIP
  • C&R
 H Mk 205 Rabbit IgG
GLTSCR1L (E9K6F) Rabbit mAb 92805 20 µl
  • WB
  • IP
 H M R Mk 140 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Rab Goat 

Product 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.

Specificity / Sensitivity

Each antibody in the Non-Canonical BAF Complex Antibody Sampler Kit detects endogenous levels of its target protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro576 of human BRD9 protein, Pro1545 of human GLTSCR1 protein, recombinant protein specific to the amino terminus of human Brg1 protein and human GLTSCR1L protein.

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).

Limited Uses

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