Revision 4

#59325Store 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, ChIP, ChIP-seq

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9GZV8

Entrez-Gene Id:

63978

Product Information

Product Usage Information

For optimal ChIP and 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Chromatin IP 1:100
Chromatin IP-seq 1:100

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

PRDM14 (E2J8Q) Rabbit mAb recognizes endogenous levels of total PRDM14 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PRDM14 protein.

Background

PR domain zinc finger protein 14 (PRDM14) is a likely protein lysine methyltransferase that is primarily expressed in primordial germ cells and pluripotent embryonic stem cells. It is essential for the establishment and maintenance of primordial germ cells and critical for the maintenance of pluripotency in embryonic stem cells (1-3). PRDM14 represses genes involved in the differentiation of stem cells into various cell lineages, likely via a combination of interactions with TET proteins, the polycomb repressive complex 2 (PRC2), and CBFA2T2 (3-8). In addition, overexpression of PRDM14 in combination with Jarid2 promotes induced pluripotent stem cell (iPSC) formation (9). PRDM14 protein levels are overexpressed in certain cancers, including breast, leukemia (T-ALL), and non-small cell lung cancer (NSCLC) (10-13), and PRDM14 overexpression may serve as a novel prognostic marker in NSCLC (14). Targeting PRDM14 overexpression with a siRNA-based therapy was shown to decrease liver metastasis in a murine pancreatic cancer model, suggesting potential as a therapeutic option for cancers where this protein is abnormally expressed (15).

  1. Yamaji, M. et al. (2008) Nat Genet 40, 1016-22.
  2. Chia, N.Y. et al. (2010) Nature 468, 316-20.
  3. Tsuneyoshi, N. et al. (2008) Biochem Biophys Res Commun 367, 899-905.
  4. Ma, Z. et al. (2011) Nat Struct Mol Biol 18, 120-7.
  5. Okashita, N. et al. (2014) Development 141, 269-80.
  6. Yamaji, M. et al. (2013) Cell Stem Cell 12, 368-82.
  7. Nady, N. et al. (2015) Elife 4, e10150.
  8. Tu, S. et al. (2016) Nature 534, 387-90.
  9. Iseki, H. et al. (2016) Stem Cells 34, 322-33.
  10. Moelans, C.B. et al. (2010) Mod Pathol 23, 1029-39.
  11. Nishikawa, N. et al. (2007) Cancer Res 67, 9649-57.
  12. Carofino, B.L. et al. (2013) Dis Model Mech 6, 1494-506.
  13. Liu, B. et al. (2010) Zhongguo Fei Ai Za Zhi 13, 867-72.
  14. Zhang, T. et al. (2013) Med Oncol 30, 605.
  15. Moriya, C. et al. (2017) Carcinogenesis 38, 638-648.

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

WB: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

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.
SimpleChIP is a registered 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 4
#59325

PRDM14 (E2J8Q) Rabbit mAb

Western Blotting Image 1: PRDM14 (E2J8Q) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using PRDM14 (E2J8Q) Rabbit mAb (upper) and -Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, HCT 116 and A549 cell lines are negative for PRDM14 expression.
Immunoprecipitation Image 1: PRDM14 (E2J8Q) Rabbit mAb Expand Image
Immunoprecipitation of PRDM14 from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is PRDM14 (E2J8Q) Rabbit mAb. Western blot analysis was performed using PRDM14 (E2J8Q) Rabbit mAb.
Chromatin Immunoprecipitation Image 1: PRDM14 (E2J8Q) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either PRDM14 (E2J8Q) Rabbit mAb or PRDM14 (E1D5S) XP® Rabbit mAb #83527, using SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA libraries were prepared using DNA Library Prep Kit for Illumina #56795. The figures show binding across ASXL1, a known target gene of PRDM14 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
Chromatin Immunoprecipitation Image 2: PRDM14 (E2J8Q) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either PRDM14 (E2J8Q) Rabbit mAb #59325 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 ASXL1 Upstream Primers #34755, SimpleChIP® Human Oct-4 Promoter Primers #4641, 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.