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MYST Antibody Sampler Kit
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MYST Antibody Sampler Kit #99991

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Western blot analysis of extracts from various cell lines using MYST1 (D5T3R) Rabbit mAb.
Western blot analysis of extracts from various cell lines using MYST2 (D4N3F) Rabbit mAb.
CUT&RUN was performed with HCT116 cells and MYST2 (D4N3F) 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 ME1 gene, a known target gene of MYST2 (see additional figure containing CUT&RUN-qPCR data).
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 Hep G2, KYSE-70, and A-498 cells using MYST3 (E3P5T) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). Low expression of MYST3 protein in A-498 cells is consistent with the predicted expression pattern.
Immunoprecipitation of MYST2 from MOLT4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MYST2 (D4N3F) Rabbit mAb. Western blot analysis was performed using MYST2 (D4N3F) Rabbit mAb.
CUT&RUN was performed with HCT116 cells and MYST2 (D4N3F) 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 6 (upper), including ME1 (lower), a known target gene of MYST2 (see additional figure containing CUT&RUN-qPCR data).
Immunoprecipitation of MYST3 protein from Hep G2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MYST3 (E3P5T) Rabbit mAb. Western blot analysis was performed using MYST3 (E3P5T) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using MYST2 (D4N3F) Rabbit mAb.
CUT&RUN was performed with HCT116 cells and MYST2 (D4N3F) 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 ME1 promoter primers and human PANTR1 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.
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 days followed by treatment with β-estradiol (10 nM, 45 min) and either MYST3 (E3P5T) 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® Human ESR1 Promoter Primers #9673 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 human colon carcinoma using MYST2 (D4N3F) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using MYST2 (D4N3F) Rabbit mAb.
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and MYST2 (D4N3F) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across PRCC gene. For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and MYST2 (D4N3F) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 1 (upper), including PRCC (lower).
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either MYST2 (D4N3F) 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 HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA9 Promoter Primers #14909, 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.
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To Purchase # 99991T
Cat. # Size Price Inventory
99991T		 1 Kit  (4 x 20 microliters)

Bulk & Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
MYST1 (D5T3R) Rabbit mAb 46862 20 µl
  • WB
 H M R Mk 53 Rabbit IgG
MYST2 (D4N3F) Rabbit mAb 58418 20 µl
  • WB
  • IP
  • IHC
  • ChIP
  • C&R
 H M R Mk 80 Rabbit IgG
MYST3 (E3P5T) Rabbit mAb 85460 20 µl
  • WB
  • IP
  • ChIP
 H 320 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Rab Goat 

Product Description

The MYST Antibody Sampler Kit provides an economical means of detecting MYST family members using antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the MYST Antibody Sampler Kit detects endogenous levels of its target protein. MYST1 (D5T3R) Rabbit mAb recognizes endogenous levels of total MYST1 protein. MYST2 (D4N3F) Rabbit mAb recognizes endogenous levels of total MYST2 protein. MYST3 (E3P5T) Rabbit mAb recognizes endogenous levels of total MYST3 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with recombinant proteins surrounding Val80 of human MYST1 protein and Val222 of human MYST2 protein; with a synthetic peptide corresponding to residues surrounding Pro927 of human MYST3 protein.

Background

MYST1, also known as mammalian male absent on the first (MOF) and lysine acetyltransferase 8 (KAT8), is a member of the MYST (MOZ, YBF2, SAS2, and Tip60) family of histone acetyltransferases (1,2). As the catalytic subunit of two different histone acetyltransferase complexes, MSL and NSL, MYST1 is responsible for the majority of histone H4 lysine 16 acetylation in the cell. MYST1 also acetylates p53 on lysine 120 and is important for activation of pro-apoptotic genes (1,2). As a component of the MSL complex, MYST1 associates with MSL1, MSL2L1, and MSL3L1, and specifically acetylates histone H4 on lysine 16 (3-5). As part of the NSL complex, MYST1 associates with the MLL1 histone methyltransferase complex containing MLL1/KMT2A, ASH2L, HCFC1, WDR5, and RBBP5, and shows broader acetyltransferase activity for histone H4 on lysines 5, 8, and 16 (3-5). MYST1 plays a critical role in the regulation of transcription, DNA repair, autophagy, apoptosis, and emybryonic stem cell pluripotency and differentiation (1,2,6). Loss of MYST1 leads to a global reduction in histone H4 lysine 16 acetylation, a common hallmark found in many human cancers. A reduction of MYST1 protein levels and histone H4 lysine 16 acetylation is associated with poor prognosis in breast, renal, colorectal, gastric, and ovarian cancers (1).

MYST2, also known as HBO1 and lysine acetyltransferase 7 (KAT7), is the catalytic subunit of the HBO1 acetyltransferase complex, which consists of MYST2, MYST/ESA1-associated factor 6 (MEAF6), inhibitor of growth protein 4 (ING4) or inhibitor of growth protein 5 (ING5), and one of two families of scaffold proteins (JADE-1/2/3 or BRPF1/2/3) (7,8). The substrate specificity of the HBO1 complex is determined by the associated scaffold protein. HBO1 complexes containing JADE scaffold proteins acetylate histone H4 on lysines 5, 8, and 12, while complexes containing BRPF scaffold proteins acetylate histone H3 on lysines 14 and 23 (8). In addition, the scaffold protein appears to regulate the function of the HBO1 complex. Complexes containing JADE scaffold proteins bind to origin recognition complex 1 (ORC1) and regulate licensing of DNA replication, while HBO1 complexes containing BRPF scaffold proteins regulate transcription (8-11). MYST2 is required for regulation of cell proliferation (1), adipogenesis (12), embryonic development (13), and survival of fetal liver erythroblasts (14). In addition, MYST2 is overexpressed in several human cancers, including cancers of the testis, ovary, breast, stomach, esophagus, and bladder (15). The MYST2 gene is amplified and protein is overexpressed in breast cancers, and overexpression of MYST2 increases anchorage-independent growth of several breast cancer cell lines (16).

MYST3, also known as monocytic leukemia zinc finger protein (MOZ) and lysine acetyltransferase 6A (KAT6A), was first discovered as a fusion partner of CREBBP in acute myeloid leukemia. MYST3 contributes to Hox gene expression and segment identity during development (17-20). MYST3 forms an evolutionarily conserved complex with ING5, EAF6, and BRD1 and has been shown to be a coactivator for many different transcription factors, including PU.1, NRF2, and Runx family members (21-23). MYST3 is critical in hematopoietic stem cell maintenance, where it acts synergistically with polycomb member BMI1 (24). Inhibitors of MYST3 are being investigated for therapeutic value as they can induce senescence and decrease tumor growth (25).

  1. Yang, Y. et al. (2014) Front Med 8, 79-83.
  2. Li, X. and Dou, Y. (2010) Epigenetics 5, 185-8.
  3. Dou, Y. et al. (2005) Cell 121, 873-85.
  4. Li, X. et al. (2009) Mol Cell 36, 290-301.
  5. Cai, Y. et al. (2010) J Biol Chem 285, 4268-72.
  6. Füllgrabe, J. et al. (2013) Nature 500, 468-71.
  7. Avvakumov, N. et al. (2012) Mol Cell Biol 32, 689-703.
  8. Lalonde, M.E. et al. (2013) Genes Dev 27, 2009-24.
  9. Iizuka, M. and Stillman, B. (1999) J Biol Chem 274, 23027-34.
  10. Iizuka, M. et al. (2006) Mol Cell Biol 26, 1098-108.
  11. Miotto, B. and Struhl, K. (2010) Mol Cell 37, 57-66.
  12. Johmura, Y. et al. (2008) J Biol Chem 283, 2265-74.
  13. Kueh, A.J. et al. (2011) Mol Cell Biol 31, 845-60.
  14. Mishima, Y. et al. (2011) Blood 118, 2443-53.
  15. Iizuka, M. et al. (2009) Gene 436, 108-14.
  16. Hu, X. et al. (2009) Mol Cancer Res 7, 511-22.
  17. Borrow, J. et al. (1996) Nat Genet 14, 33-41.
  18. Champagne, N. et al. (2001) Oncogene 20, 404-9.
  19. Crump, J.G. et al. (2006) Development 133, 2661-9.
  20. Voss, A.K. et al. (2009) Dev Cell 17, 674-86.
  21. Katsumoto, T. et al. (2006) Genes Dev 20, 1321-30.
  22. Ohta, K. et al. (2007) Biochem J 402, 559-66.
  23. Bristow, C.A. and Shore, P. (2003) Nucleic Acids Res 31, 2735-44.
  24. Sheikh, B.N. et al. (2017) Exp Hematol 47, 83-97.e8.
  25. Baell, J.B. et al. (2018) Nature 560, 253-257.

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