Anti-Otx2 antibody - ChIP Grade
| Name | Anti-Otx2 antibody - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab21990 |
| Prices | $404.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | IHC-F ICC/IF ICC/IF ChIP WB IHC-P IP |
| Species Reactivities | Mouse, Chicken, Bovine, Human, Xenopus, Zebrafish |
| Antigen | Synthetic peptide corresponding to Human Otx2 aa 250 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH) |
| Blocking Peptide | Otx2 peptide |
| Description | Rabbit Polyclonal |
| Gene | OTX2 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
![Otx1 +2 was immunoprecipitated using 0.5mg Y79 whole cell extract, 5µg of Rabbit polyclonal to Otx1 +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, Y79 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 ab21990.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 37kDa; Otx1 +2, non specific - as present in control (lane 2); 37kDa: We are confident this was due to slight lane contamination and the band seen in the IP lane is our target of interest.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/5019_ab21990-199601-IPV024ab219901m30s.jpg)
Otx1 +2 was immunoprecipitated using 0.5mg Y79 whole cell extract, 5µg of Rabbit polyclonal to Otx1 +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, Y79 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 ab21990.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 37kDa; Otx1 +2, non specific - as present in control (lane 2); 37kDa: We are confident this was due to slight lane contamination and the band seen in the IP lane is our target of interest.
Mouse E6 embryos were sectioned and then fixed in paraformaldehyde. Antigen retrieval was performed using citric acid and the sections permeablized and blocked for 1 hour in serum. Embryos were stained with ab21990 (1/2500) diluted in 10% serum, 1% ovalbumin in PBSTween for 14 hours at 4°C. They were then washed and stained with a goat anti-rabbit biotin conjugated antibody. After using a biotinylated secondary, ABC and then DAB was used for color development.
ab21990 staining Otx2 in embryonic day 14.5 (E14.5) mouse eye by Immunohistochemistry (formalin fixed, paraffin embedded sections).The embryo was fixed in 10% neutral buffered formalin overnight at room temperature, then paraffin-embedded and sectioned at 4µm. Following deparaffinization and antigen retrieval in a rice steamer in 10mM sodium citrate (pH 6.0), the sections were blocked with 5% normal goat serum for half an hour at room temperature. ab21990 was diluted in PBS at 1/100, and incubated overnight at 4°C. The secondary was goat anti-rabbit Alexa®568, diluted in PBS with 1.5% normal goat serum. The mounting medium contained the nuclear stain DAPI (blue).
All lanes : Anti-Otx2 antibody - ChIP Grade (ab21990) at 1 µg/mlLane 1 : Y79 LysateLane 2 : Y79 Lysate with Otx2 peptide (ab24347) at 1 µg/mlLysates/proteins at 20 µg per lane.SecondaryAlexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution
Anti-Otx2 antibody - ChIP Grade (ab21990) at 1 µg/ml + OTX1 (Human) - Recombinant Protein at 0.1 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
The image shows staining of Otx2 in human embryonic stem cells differentiated into neuroectoderm. As would be expected, staining is nuclear.
Product References
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