Revision 1

#16144

Store at -20C

NSD Family Antibody Sampler Kit

1 Kit

(3 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

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
NSD1 (E2U6H) Rabbit Monoclonal Antibody5107620 µl300 kDaRabbit IgG
WHSC1 (D4Z8Q) Rabbit Monoclonal Antibody6512720 µl66, 152 kDaRabbit IgG
WHSC1L1 (D4N9N) Rabbit Monoclonal Antibody9205620 µl80-90, 180 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 NSD Family Antibody Sampler Kit provides an economical means of detecting NSD protein family members. 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 antibody.

Background

The nuclear receptor SET domain-containing (NSD) family of histone H3 lysine 36 (H3K36) methyltransferases contains the NSD1, WHSC1/NSD2, and WHSC1L1/NSD3 proteins. More specifically, these enzymes catalyze mono- and di-methylation of H3K36, marks that are typically associated with transcriptionally active regions of the genome (1,2). NSD1 is required for proper embryonic development (3), and mutations in NSD1 are believed to be major contributing factors to the overgrowth disorder, Sotos syndrome (4-6). NSD1 is also mutated or has altered expression in several types of cancer and is thought to exert tumor suppressive or promoting functions depending on cellular context (7-11). WHSC1/NSD2 haploinsufficiency is implicated in the developmental disorder, Wolf-Hirschhorn syndrome, which is characterized by delayed growth, mental retardation, and congenital heart defects (12). In addition, WHSC1 is overexpressed and associated with poor prognosis in a large variety of human cancers, including neuroblastoma and ovarian, hepatocellular, endometrial, and colorectal carcinoma (13-17).  WHSC1L1/NSD3 can function as an oncogene or a tumor suppressor protein and has been shown to regulate expression of several genes associated with cell cycle (18). Amplification and/or increased expression of WHSC1L1/NSD3 in breast, lung, and liver cancer increases growth and survival, and is associated with poor prognosis (18-21).

Background References

  1. Wagner, E.J. and Carpenter, P.B. (2012) Nat Rev Mol Cell Biol 13, 115-26.
  2. Bennett, R.L. et al. (2017) Cold Spring Harb Perspect Med 7, a026708. doi: 10.1101/cshperspect.a026708.
  3. Rayasam, G.V. et al. (2003) EMBO J 22, 3153-63.
  4. Kurotaki, N. et al. (2002) Nat Genet 30, 365-6.
  5. Douglas, J. et al. (2003) Am J Hum Genet 72, 132-43.
  6. Rio, M. et al. (2003) J Med Genet 40, 436-40.
  7. Bianco-Miotto, T. et al. (2010) Cancer Epidemiol Biomarkers Prev 19, 2611-22.
  8. Zhang, S. et al. (2019) J Exp Clin Cancer Res 38, 467.
  9. Cancer Genome Atlas Network collaborators (2015) Nature 517, 576-82.
  10. Berdasco, M. et al. (2009) Proc Natl Acad Sci USA 106, 21830-5.
  11. Krossa, I. et al. (2022) Cancers (Basel) 14, 4865. doi: 10.3390/cancers14194865.
  12. Stec, I. et al. (1998) Hum Mol Genet 7, 1071-82.
  13. Hudlebusch, H.R. et al. (2011) Clin Cancer Res 17, 2919-33.
  14. Yang, S. et al. (2013) Biomarkers 18, 257-63.
  15. Zhou, P. et al. (2013) Pathol Oncol Res 19, 303-9.
  16. Xiao, M. et al. (2013) J Surg Oncol 107, 428-32.
  17. Hudlebusch, H.R. et al. (2011) Cancer Res 71, 4226-35.
  18. Zhou, Z. et al. (2010) Biochem Biophys Res Commun 398, 565-70.
  19. Angrand, P.O. et al. (2001) Genomics 74, 79-88.
  20. Yang, Z.Q. et al. (2010) Cancer Res 70, 8487-97.
  21. Kang, D. et al. (2013) Genes Chromosomes Cancer 52, 126-39.

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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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

#16144

NSD Family Antibody Sampler Kit

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using NSD1 (E2U6H) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). Negative expression of NSD1 protein in Hep G2 cells is consistent with the predicted expression pattern.
undefined Image 1: NSD Family Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either NSD1 (E2U6H) 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 human BMP4 promoter primers, human ZFP36L1 upstream primers, SimpleChIP® Human GAPDH Intron 2 Primers #4478, and human GAPDH 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 2: NSD Family Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using WHSC1 (D4Z8Q) Rabbit mAb.
undefined Image 3: NSD Family 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

#16144

NSD Family Antibody Sampler Kit

Cell Signaling Technology Logo
CUT&RUN was performed with HeLa cells treated with TNFa (30ng/mL, 1hr) and WHSC1 (D4Z8Q) 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 ARID1A gene.
undefined Image 4: NSD Family 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 5: NSD Family Antibody Sampler Kit
Western blot analysis of extracts from A549 and COS-7 cell lines using WHSC1L1 (D4N9N) Rabbit mAb. As expected, this antibody detects both the long and short isoforms of WHSC1L1.
undefined Image 6: NSD Family 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

#16144

NSD Family Antibody Sampler Kit

Cell Signaling Technology Logo
Immunoprecipitation of WHSC1 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 WHSC1 (D4Z8Q) Rabbit mAb. Western blot analysis was performed using WHSC1 (D4Z8Q) Rabbit mAb.
undefined Image 7: NSD Family Antibody Sampler Kit
CUT&RUN was performed with HeLa cells treated with TNFa (30ng/mL, 1hr) and WHSC1 (D4Z8Q) 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 ARID1A gene (lower).
undefined Image 8: NSD Family Antibody Sampler Kit
Immunoprecipitation of WHSC1L1 from A549 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is WHSC1L1 (D4N9N) Rabbit mAb. Western blot analysis was performed using WHSC1L1 (D4N9N) Rabbit mAb.
undefined Image 9: NSD Family 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

#16144

NSD Family Antibody Sampler Kit

Cell Signaling Technology Logo
CUT&RUN was performed with HeLa cells treated with human TNF-α #8902 (30 ng/ml, 1 hr) and either WHSC1 (D4Z8Q) 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 IκBα Promoter Primers #5552, human IL-8 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.
undefined Image 10: NSD Family Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded MOLT-4 cell pellet (left, high-expressing) or HT29 cell pellet (right, low-expressing) using WHSC1L1 (D4N9N) Rabbit mAb.
undefined Image 11: NSD Family Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded mouse liver using WHSC1L1 (D4N9N) Rabbit mAb.
undefined Image 12: NSD Family 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

#16144

NSD Family Antibody Sampler Kit

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human small intestine using WHSC1L1 (D4N9N) Rabbit mAb.
undefined Image 13: NSD Family Antibody Sampler Kit
Flow cytometric analysis of HT-29 cells (blue) and MOLT-4 cells (green) using WHSC1L1 (D4N9N) Rabbit mAb (solid lines) or a concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 14: NSD Family 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.