- 询价
- 05-806
- 2025年07月16日
- Flow Cytometry;Immunofluorescence
- Mouse
- Human
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- 详细信息
- 文献和实验
- 技术资料
- 应用范围：
Flow Cytometry;Immunofluorescence
- 宿主：
Mouse
- 适应物种：
Human
- 库存：
大量
- 抗原来源：
0
- 是否单克隆：
1
- 规格：
100 μg
IF: positive chromosome immunostaining for mitotic A431 and HeLa cells fixed with 95% ethanol and 5% acetic acid and permeabilized with 0.1% Triton X-100
WB: Acid extract from untreated (Lane 1) and colcemid treated (Lane 2) HeLa cells, along with recombinant Histone H3 (Lane 3) were probed with anti-phospho-Histone H3 (Ser10) (0.1μg/ml).
Beadlyte® Histone-Peptide Specificity Assay
Representative data from a previous lot. 1:1,000-1:81,000 dilutions of anti-phospho-Histone H3 (Ser10) were incubated with a cocktail of microspheres conjugated to histone H3 peptides with the following modifications:
1. phospho-serine 10
2. phospho-serine 28
3. phospho-threonine 11
4. No modifications, containing aa 1-20
Unbound antibody was removed by filtration. The peptide-antibody complexes were incubated with a biotin-conjugated anti-mouse secondary antibody followed by incubation with a phycoerythrinstreptavidin conjugate. Fluorescence was read on a
Luminex® 100™ instrument. Median fluorescence intensity (MFI) is plotted.
Peptide Inhibition Analysis
Representative blot from a previous lot. HeLa acid extracts from colcemid treated cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-phospho-
Histone H3 (Ser10) alone (Lane 1) or the antibody pre-absorbed with 10 μM of histone H3 peptides with the following modifications:
Lane 2: phospho-serine 10
Lane 3: phospho-serine 28
Lane 4: no modification
A 1:10,000 dilution of primary antibody was used. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | BD, FC, IC, IF, PIA, WB | Mouse | Purified | Monoclonal Antibody |
- H3 histone family, member T
- histone 3, H3
- histone cluster 3, H3
- Bead-based Assays
- Flow Cytometry
- Immunocytochemistry
- Immunofluorescence
- Peptide Inhibition Assay
- Western Blotting
This antibody has been reported by an independent laboratory to detect phosphorylated histone H3 using flow cytometry.
Beadlyte® Histone-Peptide Specificity Assay:
1:1,000 to 1:81,000 dilutions of a previous lot were incubated with histone H3 peptides containing various modifications conjugated to Luminex® microspheres. (Figure B). Only the peptide containing phospho-serine 10 was detected.
Immunocytochemistry:
0.2 μg/mL of a previous lot showed positive chromosome immunostaining for mitotic A431 and HeLa cells fixed with 95% ethanol and 5% acetic acid and permeabilized with 0.1% Triton® X-100.
Peptide Inhibition Analysis:
Detection of histone H3 in immunoblots of colcemid treated HeLa acid extracts by anti-phospho-Histone H3 (Ser10) was diminished by 10 μM of histone H3 peptide containing phospho-serine 10, but not by peptides containing phospho-serine 28 or an unmodified histone H3 sequence (Figure C).
Immunofluorescence:
Broad species cross-reactivity is expected.
Handling Recommendations:
Upon receipt, 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. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
- HIST3H3
- H3FT
- MGC126886
- H3t
- MGC126888
- H3T
- H3/g
- H3.4
- H3/t
Western Blot Analysis:
0.05-0.5 μg/mL of this lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid (Catalog # 17-306), but not unmodified recombinant Histone H3 (Catalog # 14-494) (Figure A).
Product Family Information
Histone H3Millipore’s Anti-Histone H3 Antibody demonstrates specificity against Histone H3. See below for data, references and related products for Histone H3. All Millipore antibodies are based on the expertise of Upstate & Chemicon. |
更多产品技术资讯，请访问密理博中国博客：http://blog.milliporechina.com
A proprietary immunogen based on a peptide sequence containing phospho-serine corresponding to residue 10 of human histone H3 详细描述见链接：http://www.millipore.com/catalogue/item/05-806
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文献和实验at 10, 25, and 50 μM). At 24 h post-treatment, celllysates were obtained; SDS-PAGE and Western blotting were performed as described in Section 3, Methods. AR-A014418dose-dependent decreases of phospho-β-catenin (Ser33/37) and phospho-glycogen synthase
Competing Chromosomal Proteins from Drosophila Polytene Chromosomes Using Modified Histone Peptides
to chromatin, by its histone tail interaction or by other interactions, then you would not expect to see loss of binding upon peptide competition. A good positive control for the assay is to use H3K9 dimethylated peptide to compete HP1 from the pericentric
v468. chapter 4 WNT 细胞通路 检测细胞GSK3活性与表达水平
solution contains 100 mMMgCl 2 , 1 mM un-labelled ATP, and approximately 30 MBq/mL [g- 32 P]ATP. 5. Phospho-glycogen synthase 2 (pGS2) peptide is available fromUpstate (#12-241) and the pGS2 10× stock solution contains200 μM pGS2 in kinase buffer
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