H3k9me2

Manufactured by Santa Cruz Biotechnology

Sourced in United States

H3K9me2 is an antibody that recognizes the di-methylation of lysine 9 on the histone H3 protein. This epigenetic modification is associated with transcriptional repression and heterochromatin formation.

Automatically generated - may contain errors

Lab products found in correlation

4 protocols using h3k9me2

Assessing Epigenetic Regulation of NF-κB

Check if the same lab product or an alternative is used in the 5 most similar protocols

To assess the binding of SirT1, H3K9me2 and H4K16ace around the NF-κB binding site of the TNF-α proximal promoter in the SirT1-treated tolerant THP-1 cells, ChIP assays (Upstate Biotechnology, Inc., Lake Placid, NY, USA) were performed with the following modifications. Proteins from 5×10⁶ cells in each sample were cross-linked to DNA using 1% formaldehyde for 10 min at room temperature. Each sample with sheared chromatin was divided into two sample groups, providing an ‘input’ sample, which was not incubated with any antibodies. The other sample was incubated with antibodies specific for SirT1 (cat. no. sc-135792; 1:500; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), H3K9me2 (cat. no. 6814-25; 1:600; BioVision, Inc., Milpitas, CA, USA), H4K16ace (cat. no. sc-8662; 1:500; Santa Cruz Biotechnology, Inc.), and IgG (1:800; cat. no. sc-2027; Santa Cruz Biotechnology, Inc.) for the negative control at 4°C overnight.

Chen G.D., Yu W.D, & Chen X.P. (2016). SirT1 activator represses the transcription of TNF-α in THP-1 cells of a sepsis model via deacetylation of H4K16. Molecular Medicine Reports, 14(6), 5544-5550.

+ Open protocol

+ Expand

Antibodies Used for Chromatin Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The antibodies used in this study were purchased from the following sources: Millipore (Billerica, MA): ASYM25 (09-814), HP1 (MAB3448), histone H3 (06-755), H3ac (06-599), H3K4me3 (07-473), H3K9me2 (07-441), H3K9me3 (07-442), H3K27me3 (07-449), H4 (05-858), H4ac (06-598), Prmt1 (07-404) and Sox2 (AB5603); Santa Cruz Biotech (Santa Cruz, CA): actin (sc-47778), c-Myc (sc-40, sc-789), GAPDH (sc-166574), Gata4 (sc-9053), Gata6 (sc-9055), GST (sc-138), mSin3a (sc-994), Oct4 (sc-5279) and p300 (sc-32244); Cell Signaling Technology (Danvers, MA): HDAC1 (2062), HDAC2 (2545s) and KLF4 (4038); Abcam (Cambridge, UK): H3R17me2 (ab412); Sigma (St. Louis, MO): FLAG (F3165); MBL (Japan): FLAG (PM020); Active Motif (Carlsbad, CA): H4R3me2a (39705); R&D Systems (Minneapolis, MN): KLF4 (AF3158); and Bethyl Laboratories (Montgomery, TX): Nanog (A300-397A).
A specific monoclonal antibody against Klf4-R396me2a was generated by Absea Biotechnology Ltd (Beijing, China) using synthesized peptide CGRRSWPRKRTATHT, corresponding to residues aa 387–402 of Klf4, as an antigen.

Zuo Z.Y., Yang G.H., Wang H.Y., Liu S.Y., Zhang Y.J., Cai Y., Chen F., Dai H., Xiao Y., Cheng M.B., Huang Y, & Zhang Y. (2022). Klf4 methylated by Prmt1 restrains the commitment of primitive endoderm. Nucleic Acids Research, 50(4), 2005-2018.

+ Open protocol

+ Expand

Western Blot Analysis of Epigenetic Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

20 μg of total cell lysates were subjected to a 10% polyacrylamide sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and blots were transferred onto polyvinylidene difluoride (PVDF) membranes. The membranes with the blots were then incubated with 5% non-fat milk in PBS with Tween-20 for 1 h to prevent non-specific binding before being incubated overnight at 4 °C in specific primary antibodies against HDAC1, HDAC7, HDAC8, E-cadherin, Vimentin, Ac-H3, Ac-H4, H3K4me2, H3K9me2 and Histone H3 (Santa Cruz Biotechnology, CA, USA) followed by incubation in peroxidase - conjugated secondary antibody at room temperature for 1 h, washed with PBST three times, then the protein signals were observed using the UVP BioSpectrum system (Analytic Jena Company).

Cai M.H., Xu X.G., Yan S.L., Sun Z., Ying Y., Wang B.K, & Tu Y.X. (2018). Depletion of HDAC1, 7 and 8 by Histone Deacetylase Inhibition Confers Elimination of Pancreatic Cancer Stem Cells in Combination with Gemcitabine. Scientific Reports, 8, 1621.

+ Open protocol

+ Expand

Antibody Sourcing for Epigenetic Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies were purchased from Diagenode for 5mC antibody (3D33), MBD1 N-terminal (#078-050), MeCP2 (#052-050), H3K27ac (C15410174); from Active motif for DNMT1 (#39-906), MBD2 (#39-548); from Abcam for RNAPII pS2 (ab5095), RNAPII pS5 (ab5131), histone H3 (Ab1791), H3K9me3 (ab8898); from Euromedex for HP1a (2HP-2G9-AS), BRG1 (2SN-2E12-AS); from Millipore for HP1γ (42s2, 05-690), H3K9me2 (07–441); from Santa-Cruz for Sam68 (7-1, sc-1238), ASF/SF2 (sc-33652), U2AF65 (MC3, sc-53942), RNAPII (N20, sc-899), N-cadherin (CDH2, sc-7939); from Cell Signaling for SUV39H1 (D11B6, #8729); from SIGMA for EHMT2 (HPA050550); from Novusbio for N-terminal-Cas9/dCas9 (79A-3A3); from ThermoFisher for ESRP1 (PA5-25833), E-cadherin (CDH1, PA5-32178), Vimentin (PA5-27231), CD44v5 (VFF-8, MA5-16967); and from eBioscience for CD44v7-v8 (VFF-17, BMS118). Hermes3 ascite directed against CD44 constant exons was a kind gift from Larry Sherman and Peter Herrlich (University of Karlsruhe, Germany).

Batsché E., Yi J., Mauger O., Kornobis E., Hopkins B., Hanmer-Lloyd C, & Muchardt C. (2021). CD44 alternative splicing senses intragenic DNA methylation in tumors via direct and indirect mechanisms. Nucleic Acids Research, 49(11), 6213-6237.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!