Log In Sign up
The largest database of trusted experimental protocols

H3k36me2

Manufactured by Merck Group
Visit Supplier

H3K36me2 is a modified histone H3 protein that contains a dimethylation at the lysine 36 residue. This modification plays a role in regulating gene expression and chromatin structure. The H3K36me2 antibody can be used to detect and analyze this specific histone modification in various laboratory applications.

Automatically generated - may contain errors

Lab products found in correlation

H3k36me3, by Abcam (3 mentions) H3k27me3, by Merck Group (3 mentions) H3k4me2, by Merck Group (3 mentions) H3k27me2, by Merck Group (3 mentions) H3k4me3, by Merck Group (3 mentions) H3k9me2, by Merck Group (2 mentions) H3k27me1, by Merck Group (2 mentions) H3k9me2, by Abcam (2 mentions) H3k9me3, by Merck Group (2 mentions) H3k36me1, by Abcam (1 mentions) Hdac2, by Merck Group (1 mentions) C myc, by Abcam (1 mentions) Horseradish peroxidase conjugated donkey anti rabbit igg, by GE Healthcare (1 mentions) Nuclear complex co ip kit, by Active Motif (1 mentions) Flag tag (flag), by Merck Group (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Apc cd235a, by Thermo Fisher Scientific (1 mentions) Parp 1, by Santa Cruz Biotechnology (1 mentions) H3 sc 8654, by Santa Cruz Biotechnology (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Anti-H3K36me2 (5 citations)

9 protocols using h3k36me2

1

ChIP-seq Analysis of H3K27me3 and H3K36me2

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
CD8+ splenic T cells from wild-type and cTKO mice were isolated by flow cytometry. ChIP against H3K27me3 (1 μg; Millipore, 07–449) and H3K36me2 (1 μg; Millipore, 07–274) and sequencing library preparation was performed exactly as previously described26 (link). Adapters were trimmed using BBTools28 (link). Alignment to mm10 was performed using Bowtie221 (link) and duplicate reads were marked using Picard and removed using SAMtools. Signal normalization between samples was performed using THOR30 (link). ChIP-seq signal intensity was calculated for chromatin states using GRanges and custom R scripts (Supplementary File 1).
Willcockson M.A., Healton S.E., Weiss C.N., Bartholdy B.A., Botbol Y., Mishra L.N., Sidhwani D.S., Wilson T.J., Pinto H.B., Maron M.I., Skalina K.A., Toro L.N., Zhao J., Lee C.H., Hou H., Yusufova N., Meydan C., Osunsade A., David Y., Cesarman E., Melnick A.M., Sidoli S., Garcia B.A., Edelmann W., Macian F, & Skoultchi A.I. (2020). H1 histones control the epigenetic landscape by local chromatin compaction. Nature, 589(7841), 293-298.
+ Open protocol
+ Expand
2

Whole-Cell Yeast Extract Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole-cell extracts for yeast were prepared according to protocol published elsewhere25 (link). Briefly, 5 ml yeast cultures grown to the mid-log phase were collected, washed once with water and resuspended in 400 μl of 2 M NaOH with 8% β-mercaptoethanol. The suspension was incubated on ice for 5 min, followed by a 10 min spin at 16,000 g. The supernatant was removed and the pellet was resuspended in 400 μl of Buffer A (40 mM HEPES-KOH pH 7.5, 350 mM NaCl. 0.1% Tween-20, 10% glycerol and protease inhibitors) incubated in ice for 10 min and harvested as above. The resultant pellet was resuspended in 2 × SDS loading dye and separated on a 15% SDS–PAGE gel. The gel was transferred to an activated PVDF membrane using the wet-transfer method. Antibodies used for chromatin immunoprecipitation (ChIP) and western blotting are H3 (Abcam, #1791), H3K36me3 (Abcam, #9050) H3K36me2 (Millipore, #07–369) and H3K36me1 (Abcam, #9048). All the antibodies were used at a dilution of 1:1,000 for the western blot. Original uncropped blots of all western blots used for the figures are shown in Supplementary Fig. 9.
Venkatesh S., Li H., Gogol M.M, & Workman J.L. (2016). Selective suppression of antisense transcription by Set2-mediated H3K36 methylation. Nature Communications, 7, 13610.
+ Open protocol
+ Expand
3

Nuclear Protein Extraction and Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
Nuclear proteins were extracted using the Nuclear Complex Co-IP Kit (Active Motif). Proteins were electrophoretically separated, blotted and detected using enhanced chemiluminescence. Primary antibodies used were: H3K36me2 (Millipore 07-369), H3K27me3 (Millipore 07-449), MMSET [12] (link), c-MYC (Abcam ab32072), HDAC2 (Millipore 05-814) and pan-H4 (Abcam ab7311). The secondary antibody used was horseradish peroxidase-conjugated donkey anti-rabbit IgG (GE Healthcare Life Sciences).
Popovic R., Martinez-Garcia E., Giannopoulou E.G., Zhang Q., Zhang Q., Ezponda T., Shah M.Y., Zheng Y., Will C.M., Small E.C., Hua Y., Bulic M., Jiang Y., Carrara M., Calogero R.A., Kath W.L., Kelleher N.L., Wang J.P., Elemento O, & Licht J.D. (2014). Histone Methyltransferase MMSET/NSD2 Alters EZH2 Binding and Reprograms the Myeloma Epigenome through Global and Focal Changes in H3K36 and H3K27 Methylation. PLoS Genetics, 10(9), e1004566.
+ Open protocol
+ Expand
4

Antibody Panel for Epigenetic Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies against H3K36me2 (Millipore, Billerica, MA; 07-274, dilution (1:2000)), BRCA1 (07-434 dilution (1:1000)), Flag (Sigma, St. Louis, MO; F3165, dilution into 1 ng/ml)), NSD2 (EpiCypher, Durham, NC; 13-0002 dilution (1:1000)), MMSET/NDS2 (Abcam, ab75359 dilution (1:1000)), APC-CD235A (eBioscience, Waltham, MA;17-9987-42), FK2 (Enzo Life Sciences, Farmingdale, NY; BML-pw-8810-0100, dilution (1:1000)), β-actin (sc-47778 dilution (1:1000)), H3 (sc-8654, dilution (1:1000)), PARP1 (sc-56197, dilution (1:1000)), and tubulin (sc-9103, dilution (1:1000)) (all from Santa Cruz Biotechnology, Dallas, TX) were employed.
Park J.W., Kang J.Y., Hahm J.Y., Kim H.J, & Seo S.B. (2020). Proteosomal degradation of NSD2 by BRCA1 promotes leukemia cell differentiation. Communications Biology, 3, 462.
+ Open protocol
+ Expand
5

Histone Modification Profiling Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies against GAPDH (CSB-PA00025A0Rb; Cus Ab); AURKA (sc-373856), YY1 (sc-7341), p21 (sc-397), and H3 (sc-8654) (Santa Cruz Biotechnology); H3K4me2 (07-030), H3K9me2 (07-441), H3K27me2 (07-452), and H3K36me2 (07-274) (Millipore); and H3S10ph (9701S) (Cell Signaling Technology) were used for western blot and ChIP analyses.
Park J.W., Cho H., Oh H., Kim J.Y, & Seo S.B. (2018). AURKA Suppresses Leukemic THP-1 Cell Differentiation through Inhibition of the KDM6B Pathway. Molecules and Cells, 41(5), 444-453.
+ Open protocol
+ Expand
6

Mammary Cell Culture and Epigenetic Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human primary mammary cells were purchased from Chi Scientific and cultured in DMEM/F12 (10 % FBS, with addition of insulin, hydrocortisone, and EGF) at 37 °C with 5 % CO2. MCF7 was purchased from Cell Bank of Chinese Academy and cultured in DMEM with FBS, insulin (10 μg/mL), sodium pyruvate (Invitrogen), and nonessential amino acid (Invitrogen); T47D is a gift from Dr. Yong-Feng Shang of Tianjin Medical University and cultured in RPMI1640 with FBS and insulin. Both the primary cells and cancer cell lines were sub-cultured 1:4 on reaching confluence; each passage was considered two PD.
Antibodies were purchased from the indicated companies: H3K9ac, hTERT, KRT14, and ACTG (Epitomics); KRT18 (ProteinTech), JMJD1B, JMJD2B, JMJD2A, H4K20me3, H3K79me2, E-cadherin, and RAS (CST); H3K4me1, H3K4me3, H3K27me3, H3K27me1, H3K27me2, and H3K36me2 (Millipore); H3, H3K9me1, H3K4me2, and H3K36me3 (Abcam); H3K9me2, H3K9me3, GAPDH, EHMT2, SUV39H1, CBX5, DNMT3A, DNMT3B, DNMT1, and KDM1A (Abclonal); KDM3A/JMJD1A (Abclonal for western and Millipore for immunostaining). The information of primers and siRNAs are listed in Additional file 2: Table S16.
Zhao Q.Y., Lei P.J., Zhang X., Zheng J.Y., Wang H.Y., Zhao J., Li Y.M., Ye M., Li L., Wei G, & Wu M. (2016). Global histone modification profiling reveals the epigenomic dynamics during malignant transformation in a four-stage breast cancer model. Clinical Epigenetics, 8, 34.
+ Open protocol
+ Expand
7

Visualizing Histone Methylation Patterns

Check if the same lab product or an alternative is used in the 5 most similar protocols
Nuclei transfected with JMJ13-GFP or JMJ13H293A/E295A-GFP mutants were visualized by observing the GFP signal under a fluorescence microscope. Immunolabeling was performed by using histone methylation-specific antibodies (H3K27me3, Millipore 07-449; H3K27me2, Millipore 07-452; H3K27me1, Millipore 07-448; H3K4me3, Millipore 07-473; H3K4me2, Millipore 07-030; H3K4me1, Millipore 07-436; H3K9me3, Millipore 07-442; H3K9me2, Millipore 07-441; H3K9me1, Millipore 07-450; H3K36me3, Abcam ab9050; H3K36me2, Millipore 07-274; H3K36me1, Millipore 07-548). All these antibodies were diluted by 1:100. Alexa Fluor 555 or 488-conjugated goat anti-rabbit (1:500, Invitrogen) were used as secondary antibodies to determine the specific lysine modification site in vivo. After incubation with secondary antibody, nuclei on the slide were mounted by one drop of VECTASHIELD Mounting Medium with 4′,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories), then photographed under a fluorescent microscope (Olympus BX51). ImageJ (National Institutes of Health) was used for quantification of the immunolabeled nuclei.
Zheng S., Hu H., Ren H., Yang Z., Qiu Q., Qi W., Liu X., Chen X., Cui X., Li S., Zhou B., Sun D., Cao X, & Du J. (2019). The Arabidopsis H3K27me3 demethylase JUMONJI 13 is a temperature and photoperiod dependent flowering repressor. Nature Communications, 10, 1303.
+ Open protocol
+ Expand
8

Drosophila Experiments: Developmental and Genetic Analyses

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fly experiments were conducted as previously described (Ferguson and Martinez-Agosto 2014 (link)). Fly stocks were maintained at 25°C on standard food containing 0.7% agar, 5% glucose and 7% dry yeast. Canton-S was used as the wild type (wt) strain for all performed experiments. The strains apterous-gal 4(II)/Cyo, GFP, UAS-Mes-4/NSD (BL 22044, BL 15384), and UAS-IRwt were obtained from the Bloomington Drosophila Stock Center. We generated a UAS-Mes-4/NSD; UAS-IRwt stock using standard methods.
Brightfield images of third instar larvae and adult flies were obtained using a Leica light microscope. Larvae images were obtained at a magnification of 2.0X, adult flies at 1.6X and adult wings at 3.2X. Third instar larvae tissues were dissected then fixed at room temperature in 3.7% formaldehyde. Tissues were washed in 0.4% Triton X-100 in primary antibody overnight at 4°C, washed, stained in secondary antibody for 2 hours, mounted using Vectashield (Vector Laboratories) and imaged using a Zeiss LSM5. Antibodies used include: Cleaved Caspase-3 (Cell Signaling; 1:50), Phospho Histone H3 (Cell signaling; 1:50), Gigas (DHSB, 1:20), and H3K36me2 (EMDMillipore, 1:100).
Quintero-Rivera F., Eno C.C., Sutanto C., Jones K.L., Nowaczyk M.J., Wong D., Earl D., Mirzaa G., Beck A, & Martinez-Agosto J.A. (2021). 5q35 duplication presents with psychiatric and undergrowth phenotypes mediated by NSD1 overexpression and mTOR signaling downregulation. Human genetics, 140(4), 681-690.
+ Open protocol
+ Expand
9

ChIP assay for histone modifications

Check if the same lab product or an alternative is used in the 5 most similar protocols
ChIP experiments were performed as described previously with minor modifications (Saleh et al., 2008 (link)). Leaf tissues (1.5 g) from 3-week-old plants were fixed with 1% (v/v) formaldehyde for 40 min at room temperature, and the chromatin samples were sonicated to yield 200–1,000-bp fragments. After pre-clearing of the chromatin samples with salmon sperm DNA/protein A agarose beads (EMD Millipore), immunoprecipitations were carried out with the appropriate antibodies to histone lysine methylation and reverse cross-linking overnight at 65°C. Immunoprecipitated DNA samples were purified using a silica membrane column (MACHEREY-NAGEL Inc.) and eluted in 60-μL elution buffer. For qPCR, 2 μL of DNA was amplified using SYBR Green Supermix (Bio-Rad) with specific primers, as listed in Supplemental Data Set S8. The data are presented as percentage of input values. The antibodies used for the ChIP experiments were: H3K4me2 (07-030, EMD Millipore), H3K4me3 (07-473, EMD Millipore), H3K9me2 (ab1220, Abcam), H3K9me3 (07-442, EMD Millipore), H3K36me2 (07-369-I, EMD Millipore), H3K36me3 (ab9050, Abcam), and IgG (sc‐2027, Santa Cruz) as a negative control.
Lee S., Fu F., Liao C.J., Mewa D.B., Adeyanju A., Ejeta G., Lisch D, & Mengiste T. (2022). Broad-spectrum fungal resistance in sorghum is conferred through the complex regulation of an immune receptor gene embedded in a natural antisense transcript. The Plant Cell, 34(5), 1641-1665.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!