Anti-Histone H1.5 antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab18208
• Prices: $395.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ChIP WB IHC-P ICC/IF ICC/IF
• Species Reactivities: Human
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H1
• Blocking Peptide: Human Histone H1.5 peptide
• Description: Rabbit Polyclonal
• Gene: HIST1H1B
• Conjugate: Unconjugated

Product images

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab18208 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

All lanes : Anti-Histone H1.5 antibody - ChIP Grade (ab18208) at 0.5 µg/mlLane 1 : Recombinant histone H1Lane 2 : Recombinant histone H1.1Lane 3 : Recombinant histone H1.2Lane 4 : Recombinant histone H1.3Lane 5 : Recombinant histone H1.4Lane 6 : Recombinant histone H1.5

ICC/IF image of ab18208 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18208, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879 Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

IHC image of ab18208 staining in human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18208, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Product References

Proteomic analysis of phosphorylated nuclear proteins underscores novel roles for - Proteomic analysis of phosphorylated nuclear proteins underscores novel roles for

Laserna EJ, Valero ML, Sanz L, del Pino MM, Calvete JJ, Barettino D. Mol Endocrinol. 2009 Nov;23(11):1799-814.

Depletion of human histone H1 variants uncovers specific roles in gene expression - Depletion of human histone H1 variants uncovers specific roles in gene expression

Sancho M, Diani E, Beato M, Jordan A. PLoS Genet. 2008 Oct;4(10):e1000227.