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16144
NSD Family Antibody Sampler Kit
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NSD Family Antibody Sampler Kit #16144

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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.
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.
Western blot analysis of extracts from various cell lines using WHSC1 (D4Z8Q) Rabbit mAb.
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.
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 A549 and COS-7 cell lines using WHSC1L1 (D4N9N) Rabbit mAb. As expected, this antibody detects both the long and short isoforms of WHSC1L1.
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.
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).
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.
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.
Immunohistochemical analysis of paraffin-embedded MOLT-4 cell pellet (left, high-expressing) or HT29 cell pellet (right, low-expressing) using WHSC1L1 (D4N9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse liver using WHSC1L1 (D4N9N) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human small intestine using WHSC1L1 (D4N9N) Rabbit mAb.
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.
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To Purchase # 16144T
Cat. # Size Price Inventory
16144T		 1 Kit  (3 x 20 microliters)

Bulk & Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
NSD1 (E2U6H) Rabbit mAb 51076 20 µl
  • WB
  • ChIP
 H Mk 300 Rabbit IgG
WHSC1 (D4Z8Q) Rabbit mAb 65127 20 µl
  • WB
  • IP
  • C&R
 H Mk 66, 152 Rabbit IgG
WHSC1L1 (D4N9N) Rabbit mAb 92056 20 µl
  • WB
  • IP
  • IHC
  • F
 H M R Mk 80-90, 180 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Goat 

Product 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.

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).

  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.

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