| Reactivity: |
Human |
| Validated Application: |
Chromatin Immunoprecipitation, ELISA, Dot Blot, Western Blot |
| Host: |
Rabbit |
| SubClass (Isotype): |
IgG |
| Clonality: |
Polyclonal |
| Target Gene: |
H3K27me2 |
| Target Molecule: |
Peptides |
| Gene ID: |
8290 |
| Gene Symbol: |
HIST3H3 |
| Accession Number: |
NM_003493 |
| Label or Dye: |
Unconjugated |
| Target Modification State: |
Methylated |
| Purification: |
Affinity Purified |
| Concentration: |
0.35 mg⁄ml |
| Color: |
Unconjugated |
| Form: |
Frozen |
| High Throughput Compatibility: |
Manual Protocols |
| Neutralizing Antibody: |
No |
| Product Size: |
50 µg |
| Volume: |
143 µl |
| cGMP Compliance: |
non-cGMP Compliant |
| Shipping Condition: |
Dry Ice |
| Regulatory Statement: |
For Research Use Only. Not for use in diagnostic procedures. |
Store at -20°C⁄ -80°C. Avoid multiple freeze-thaw cycles. Guaranteed stable for 6 months when properly stored.
Note: Use lot number only (e.g., "K18126").
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![Titer determination.](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/49-1013_H3K27me2_F1.jpg","Titer determination.","To determine the titer, an ELISA was performed using anti-H3K27me2 crude serum, the anti-H3K27me2 antibody (Cat. no. 49-1013), and the column flow through obtained from the antibody purification step. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:4,350.");)
![Cross reactivity ...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/49-1013_H3K27me2_F2.jpg","Cross reactivity tests using the anti-H3K27me2 antibody.","A dot blot analysis was performed to test the cross reactivity of the anti-H3K27me2 (Cat. no 49-1013) with other histone H3 modifications. Other H3 modifications include mono- and trimethylation of the same lysine and mono-, di-, and trimethylation of lysine 9, 36, and 79. 100 to 0.2 pmol of peptide containing the respective histone modification was spotted on a membrane and detected with the antibody diluted 1:20,000. This figure shows a high specificity of the antibody for the modification of interest.");)
![Western blot anal...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/49-1013_H3K27me2_F3.jpg","Western blot analysis using the anti-H3K27me2 antibody.","Histone extracts (10 µg) from NB4 cells were analyzed by western blot using anti-H3K27me2 antibody (Cat. no. 49-1013), diluted 1:500 in TBS-Tween. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.");)
![ChIP results obta...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/49-1013_H3K27me2_F4.jpg","ChIP results obtained with the anti-H3K27me2 antibody.","ChIP assays were performed using human osteosarcoma (U-2OS) cells, anti-H3K27me2 antibody (Cat. no. 49-1013), and optimized PCR primer sets for qPCR. Each ChIP assay used sheared chromatin from 1.5 million cells and various amounts of anti-H3K27me2 antibody (2, 5, and 10 µg). IgG (5 µg⁄IP) was used as negative IP control. qPCR was performed with primers for the GAPDH promoter and for exon 2 of the myoglobin gene. This figure shows the recovery, expressed as a percentage of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me2 is preferably present at silent genes. ");)
