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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| M, H, R | WB, ChIP, IP, DB |
| Description | |
|---|---|
| Catalogue Number | 17-10498 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ Monomethyl-Histone H3 (Lys36) - ChIP Validated Antibody and Primer Set |
| Overview | All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+™ Monomethyl-Histone H3 (Lys36) set includes the monomethyl-Histone H3 (Lys36) antibody, a Normal Rabbit IgG, and control primers which amplify a 213 bp region of the human GAPDH coding region. The Monomethyl-Histone H3 (Lys36) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Monomethyl-Histone H3 (Lys36) associated chromatin. |
| Alternate Names |
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| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | Anti-Monomethyl-Histone H3 (Lys36) (Rabbit Polyclonal). One vial containing 75 µL 0.35 mg/mL purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH7.4),150mM NaCl, and 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C. Normal Rabbit IgG. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide. Store at -20°C. ChIP Primers, GAPDH coding D2. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH coding region. Store at -20°C. FOR: GCC ATG TAG ACC CCT TGA AGA G REV: ACT GGT TGA GCA CAG GGT ACT TTA T |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | The ChIPAb+ Monomethyl-Histone H3 (Lys36) set includes the monomethyl-Histone H3 (Lys36) antibody, a Normal Rabbit IgG & control primers which amplify a 213 bp region of the human GAPDH coding region. |
| Key Applications |
|
| Application Notes | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of Normal Rabbit IgG, or 3 µL (1 µg) Anti-Monomethyl-Histone H3 (Lys36) and the Magna ChIP™ HiSens (Cat. # 17-10460). Successful immunoprecipitation of monomethyl-Histone H3 (Lys36) associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH coding D2 as a positive locus, and GAPDH promoter as a negative locus. Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated. Please refer to the Magna ChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens(Cat. # 17-10461) protocol for experimental details. Western Blotting Analysis: HeLa acid extract and various recombinant Histone proteins were probed with Anti-monomethyl-Histone H3 (Lys36) (0.05 µg /mL). Immunoprecipitation Analysis: 10 µg from a representative lot immunoprecipitated monomethyl-Histone H3 (Lys36) from HeLa acid extract. 10 µg of this antibody immunoprecipitated monomethyl -Histone H3 (Lys36) from 500 µg of HeLa acid extract. Arrow indicates monomethyl-Histone H3 (Lys36) (~17 kDa). Dot Blot Specificity Analysis: Unmodified and various modified Histone peptides were probed with Anti-monomethyl-Histone H3 (Lys36) (0.5 µg/mL).Histone peptides with various modifications were transferred to PVDF membrane (Cat. #: 16-665) and probed with Anti-monomethyl-Histone H3 (Lys36) at (0.5 µg/mL). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to the N-terminus of human Histone H3 monomethylated at Lys36. |
| Epitope | Monomethylated Histone H3 Lys36 |
| Host | Rabbit |
| Specificity | This antibody recognizes the N-terminus of Histone H3 monomethylated at Lys36. |
| Species Reactivity |
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| Species Reactivity Note | Demonstrated to react with human, mouse and rat. Wide species cross reactivity is predicted based on sequence similarity. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq] |
| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | Function: Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Subunit structure: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Subcellular location: Nucleus, Chromosome Tissue specificity: Expressed in testicular cells. Developmental stage: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. Post-translational modification: Methylation at Lys-36 (H3K36me) is linked to gene activation. Sequence similarities: Belongs to the histone H3 family. |
| Molecular Weight | ~17 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Rabbit IgG, or 3 µL (1 µg) Anti-Monomethyl-Histone H3 (Lys36) and the Magna ChIP™ HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of monomethyl Histone H3 (Lys36) associated DNA fragments was verified by qPCR using ChIP Primers, GAPDH coding D2. Please refer to the MagnaChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) protocol for experimental details. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: 3 μL (1 μg) of antibody per chromatin immunoprecipitation (dependent upon biological context). |