Anti-Histone H3 antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab12079
• Prices: $400.00
• Sizes: 100 µg
• Host: Goat
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ChIP IP ChIP ChIP WB IHC-P
• Species Reactivities: Mouse, Chicken, Dog, Human, Yeast, Xenopus, Drosophila, Hamster, Arabidopsis thaliana, C. elegans, S. pombe
• Antigen: Synthetic peptide derived from within residues 100 to the C-terminus of Human Histone H3
• Description: Goat Polyclonal
• Gene: CELE_F58A4.3
• Conjugate: Unconjugated

Product images

IHC image of Histone H3 staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol B. 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 ab12079, 5µg/ml, for 15 mins at room temperature. A donkey anti-goat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.

Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The  ChIP was performed with 25 µg of chromatin, 2 µg of  ab12079 (blue), and 20 µl 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.

Anti-Histone H3 antibody - ChIP Grade (ab12079) at 0.5 µg/ml + HeLa Cell Histone prep at 5 µgSecondaryRabbit Anti-Goat IgG H&L (HRP) (ab6741) at 1/5000 dilutionPerformed under reducing conditions.

All lanes : Anti-Histone H3 antibody - ChIP Grade (ab12079) at 1 µg/mlLane 1 : Chicken S1:29 Cell lysate at 10 µgLane 2 : CHO Cell lysate at 10 µgLane 3 : Dog MDCK Cell lysate at 10 µgLane 4 : Mouse 3T3 Whole Cell LysateLane 5 : Xenopus embryo Cell lysate at 10 µgSecondaryAlexa Fluor 680 Rabbit anti-Goat IgG at 1/10000 dilutionPerformed under reducing conditions.

Product References

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