Revision 6

#46395

Store at -20C

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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.

Applications:
W, IP, IF-IC, FC-FP, ChIP, ChIP-seq, C&R, C&T

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
65

Source/Isotype:
Rabbit IgG

UniProt ID:
#P25490

Entrez-Gene Id:
7528

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunofluorescence (Immunocytochemistry) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:100
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50
CUT&Tag 1:50

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 antibody.

Specificity/Sensitivity

YY1 (D5D9Z) Rabbit mAb recognizes endogenous levels of total YY1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human YY1 protein. The antibody epitope has been further mapped to a peptide corresponding to residues surrounding Pro284 of human YY1 protein.

Background

YY1 (Yin Yang1) is a ubiquitously expressed transcription factor with fundamental roles in embryogenesis, differentiation, replication and proliferation. YY1 contains four zinc finger motifs of the Cys-Cys-His-His type and can activate different eukaryotic genes (such as CREB, c-myc, Histone H4, p53 and PARP-1) or repress different eukaryotic genes (such as α-actin, IFN-β and IFN-γ) as well as regulate some viral promoters (1). YY1 deficient embryos die approximately at the time of implantation, suggesting that YY1 has an essential role in embryonic development (2). YY1 is overexpressed in cancer cells such as prostate cancer and therefore may be considered a prognostic marker (1).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

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

Alexa Fluor is a registered trademark of Life Technologies Corporation.

The transfer of this product is contingent on the buyer using the purchased product solely in research conducted by the buyer (whether the buyer is an academic or for-profit entity), for Immunocytochemistry, high content screening (HCS) analysis, or flow cytometry applications. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) resale, whether or not such product or its components are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].

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

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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 6

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using YY1 (D5D9Z) Rabbit mAb.
Western Blotting Image 1: YY1 (D5D9Z) Rabbit mAb
Immunoprecipitation of YY1 from MOLT-4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is YY1 (D5D9Z) Rabbit mAb. Western blot analysis was performed using YY1 (D5D9Z) Rabbit mAb and Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127.
Immunoprecipitation Image 1: YY1 (D5D9Z) Rabbit mAb
Confocal immunofluorescent analysis of HeLa cells using YY1 (D5D9Z) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Immunofluorescence Image 1: YY1 (D5D9Z) Rabbit mAb
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 6

Cell Signaling Technology Logo
Flow cytometric analysis of MCF7 cells using YY1 (D5D9Z) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 1: YY1 (D5D9Z) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either YY1 (D5D9Z) 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® CXXC1 Exon 1 Primers #67406, FAM193B promoter primers, 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.
Chromatin Immunoprecipitation Image 1: YY1 (D5D9Z) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and YY1 (D5D9Z) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 5 (upper), including FAM193B (lower), a known target gene of YY1 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 1: YY1 (D5D9Z) Rabbit mAb
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 6

Cell Signaling Technology Logo
CUT&RUN was performed with K562 cells and YY1 (D5D9Z) 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 RBM15 gene, a known target gene of YY1 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 1: YY1 (D5D9Z) Rabbit mAb
CUT&RUN was performed with K562 cells and YY1 (D5D9Z) 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 chromosome #1 (upper), including RBM15 (lower), a known target gene of YY1 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 2: YY1 (D5D9Z) Rabbit mAb
CUT&RUN was performed with K562 cells and either YY1 (D5D9Z) 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 RBM15 promoter primers and human ADGRL2 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.
CUT & RUN Image 3: YY1 (D5D9Z) Rabbit mAb
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 6

Cell Signaling Technology Logo
CUT&Tag was performed with K-562 cells and YY1 (D5D9Z) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across the RBM15 gene.
CUT & Tag Image 1: YY1 (D5D9Z) Rabbit mAb
CUT&Tag was performed with K-562 cells and YY1 (D5D9Z) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 1 (upper), including the RBM15 gene (lower).
CUT & Tag Image 2: YY1 (D5D9Z) Rabbit mAb
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.