Revision 5

#4402Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, FC-FP, ChIP, C&R

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

82-90

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9H334

Entrez-Gene Id:

27086

Product Information

Product Usage Information

For optimal ChIP or ChIP-seq results, use 5 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.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:100
Flow Cytometry (Fixed/Permeabilized) 1:50
Chromatin IP 1:100
CUT&RUN 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.

For a carrier free (BSA and azide free) version of this product see product #85650.

Specificity / Sensitivity

FoxP1 (D35D10) XP® Rabbit mAb detects endogenous levels of total FoxP1 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu584 of human FoxP1 protein.

Background

Forkhead box (Fox) proteins are a family of evolutionarily conserved transcription factors containing a sequence known as Forkhead box or winged helix DNA binding domain (1). The human genome contains 43 Fox proteins that are divided into subfamilies. The FoxP subfamily has four members, FoxP1 - FoxP4, which are broadly expressed and play important roles in organ development, immune response and cancer pathogenesis (2-4). The FoxP subfamily has several characteristics that are atypical among Fox proteins: their Forkhead domain is located at the carboxy-terminal region and they contain motifs that promote homo- and heterodimerization. FoxP proteins usually function as transcriptional repressors (4,5).

FoxP1 plays an important role in cardiac valve and B cell development. Targeted deletion of FoxP1 in mice results in embryonic death at E14.5 with severe defects in cardiac outflow development, endocardial cushion morphogenesis, and myocyte proliferation and maturation (6). The FoxP1 gene is located on chromosome 3p14.1 and loss of heterozygosity of this region in multiple cancer types suggests a role for FoxP1 as a tumor suppressor (7). Alternatively, FoxP1 may also act as an oncogene as it is highly expressed in a variety of B cell malignancies and is frequently involved in chromosome translocation (8,9).

  1. Myatt, S.S. and Lam, E.W. (2007) Nat Rev Cancer 7, 847-59.
  2. Shu, W. et al. (2001) J Biol Chem 276, 27488-97.
  3. Lu, M.M. et al. (2002) Gene Expr Patterns 2, 223-8.
  4. Koon, H.B. et al. (2007) Expert Opin Ther Targets 11, 955-65.
  5. Li, S. et al. (2004) Mol Cell Biol 24, 809-22.
  6. Wang, B. et al. (2004) Development 131, 4477-87.
  7. Banham, A.H. et al. (2001) Cancer Res 61, 8820-9.
  8. Wlodarska, I. et al. (2005) Leukemia 19, 1299-305.
  9. Goatly, A. et al. (2008) Mod Pathol 21, 902-11.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP C&R: CUT&RUN

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 5
#4402

FoxP1 (D35D10) XP® Rabbit mAb

Western Blotting Image 1: FoxP1 (D35D10) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using FoxP1 (D35D10) XP® Rabbit mAb.
No image available
Immunohistochemistry Image 1: FoxP1 (D35D10) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using FoxP1 (D35D10) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Flow Cytometry Image 1: FoxP1 (D35D10) XP® Rabbit mAb Expand Image
Flow cytometric analysis of MOLT-4 cells using FoxP1 (D35D10) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Chromatin Immunoprecipitation Image 1: FoxP1 (D35D10) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Raji cells and either FoxP1 (D35D10) 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 BCL2 Promoter Primers #12924, SimpleChIP® Human HSP90B1 Promoter Primers #12899, 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 and RUN Image 1: FoxP1 (D35D10) XP® Rabbit mAb Expand Image
CUT&RUN was performed with Raji cells and FoxP1 (D35D10) XP® 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 USP5 gene, a known target gene of FoxP1 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 2: FoxP1 (D35D10) XP® Rabbit mAb Expand Image
CUT&RUN was performed with Raji cells and FoxP1 (D35D10) XP® 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 12 (upper), including USP5 (lower), a known target gene of FoxP1 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 3: FoxP1 (D35D10) XP® Rabbit mAb Expand Image
CUT&RUN was performed with Raji cells and either FoxP1 (D35D10) XP® 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 USP5 promoter primers and human MYOM3 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.