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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H1.2.
Our Abpromise guarantee covers the use of ab17677 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||1/1200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IP||Use a concentration of 5 µg/ml.|
|WB||1/1000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).|
Histone H1.2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H1.2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab17677.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 35kDa; Histone H1.2
This image is courtesy of Dr Albert Jordan and Monica Sancho, Center for Genomic Regulation (CRG). Recombinant H1 isoforms courtesy of Dr Nicole Happel.
Image courtesy of Human Protein Atlas
ab17677 staining histone H1.2 in human testis, showing a distinct and strong staining pattern in ductus seminiferus and leydig cells. Paraffin embedded human skin tissue was incubated with ab17677 (1/1200 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab17677 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
ICC/IF image of ab17677 stained human MCF7 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab17677, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and HepG2 cells.