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Histone H3 K27me3/phospho S28 Antibody
SKU: orb420411
Description
Images & Validation
−Item 1 of 5
| Tested Applications | ChIP, DOT, IF, IHC, Multiplex Assay, WB |
|---|---|
| Dilution range | ChIP: 2-5µg/million cells, IHC: 1:2000, IF: 1:2000, WB: 1 µg/mL |
| Reactivity | C. elegans, Human, Mouse |
| Application Notes |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Immunogen | Histone H3 [Trimethyl Lys27, p Ser28] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with synthetic trimethylated/phosphorylated peptides surrounding Lysine 27 and Serine 28 of human Histone H3. |
| Purity | Anti-Histone H3 [Trimethyl Lys27, p Ser28] was affinity purified from monospecific antiserum by immunoaffinity chromatography. This antibody reacts with human Histone H3. A BLAST analysis was used to suggest cross-reactivity with Human, mouse, and C. elegans. Predicted to react with many species including rat, chicken, Xenopus, Drosophila, and plant based on 100% sequence homology. Cross-reactivity with Histone H3 from other sources has not been determined. |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. |
|---|---|
| Form/Appearance | Liquid (sterile filtered) |
| Buffer/Preservatives | Preservative: 0.05% (w/v) Sodium Azide. Stabilizer: 30% Glycerol; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Concentration | 0.66mg/mL |
| Dry Ice Shipping | Please note: This product requires shipment on dry ice. A dry ice surcharge will apply. |
| Disclaimer | For research use only |
Alternative Names
−rabbit anti-Histone H3 trimethyl Lys18 pS28 antibody, H3.3B, H3pS28K18Me3, H3 histone, family 3A, H3.3AH3F3H3F3B, histone H3.3, MGC87783, MGC87782, H3 K27me3/pS28
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Chromatin Immunoprecipitation Rabbit Anti-Histone H3 [Trimethyl Lys27, p Ser28] Antibody. Chromatin from one million formaldehyde cross-linked HeLa cells was used with 2 ug of Anti-Histone H3 K27me3pS28 and 20 ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and SYBR green dye, then normalized to the non-precipitated input chromatin, which is equal to one.
Dot Blot of Rabbit Histone H3 [p Ser28, Trimethyl Lys27] Antibody. Lane 1: pS28/K27 unmodified. Lane 2: pS28 N-Term. Lane 3: p28 C-term. Lane 4: K27Me3. Lane 5: pS28/K27Me3. Load: 1, 10, and 100 picomoles of peptide. Primary antibody: Histone H3 [p Ser28, Trimethyl Lys27] antibody at 1 µg/ml for 45 min at 4°C. Secondary antibody: Dylight™488 rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C.
Immunofluorescence of Rabbit Anti-Histone H3 [p Ser28, Trimethyl Lys27] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H3 [p Ser28, Trimethyl Lys27] antibody at a 1:2000 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10000 for 45 min at RT. Localization: Histone H3 [p Ser28, Trimethyl Lys27] is nuclear and chromosomal. Staining: Histone H3 [p Ser28, Trimethyl Lys27] is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 550 (red).
Immunofluorescence of Rabbit Anti-Histone H3 [p Ser28, Trimethyl Lys27] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H3 [p Ser28, Trimethyl Lys27] antibody at a 1:2000 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10000 for 45 min at RT. Localization: Histone H3 [p Ser28, Trimethyl Lys27] is nuclear and chromosomal. Staining: Histone H3 [p Ser28, Trimethyl Lys27] is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 550 (red).
Western Blot of Rabbit Anti-Histone H3 [Trimethyl Lys27, p Ser28] Antibody. Lane 1: HeLa histone prep. Lane 2: NIH-3T3 histone prep. Lane 3: C. elegans embryo cell lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [Trimethyl Lys27, p Ser28] at 1 µg/ml for overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: ~15 kDa. Other band(s): None.
Quick Database Links
UniProt
RefSeq (Protein):NP_001005464
UniProt Details
− No UniProt data available
NCBI Reference Sequences
−Associated Accession Numbers
Curated reference sequences for the gene transcript and protein product| Protein | NP_001005464 |
|---|
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Protocol Information
WB
Western Blot (IB, immunoblot)
IHC
Immunohistochemistry
IF
Immunofluorescence
DOT
Dot Blot
ChIP
Chromatin Immunoprecipitation
Histone H3 K27me3/phospho S28 Antibody (orb420411)
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