Anti-DDX58 antibody
| Name | Anti-DDX58 antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab65588 |
| Prices | $390.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | WB ICC/IF ICC/IF IP |
| Species Reactivities | Human |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 900 to the C-terminus of Human DDX58 |
| Description | Rabbit Polyclonal |
| Gene | DDX58 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
All lanes : Anti-DDX58 antibody (ab65588) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : HEK293 Human embryonic kidney cell line Whole Cell LysateLane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell LysateLane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
ICC/IF image of ab65588 stained MCF7 cells. The cells were 4% PFA fixed (10 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 (ab65588, 5µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, Hek293 and HepG2 cells fixed in PFA and MCF7 cells fixed in 100% methanol, all at 5ug/ml.
![DDX58 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to DDX58 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab65588.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 110kDa; DDX58, non-specific band: 60kDa, due to background seen in No ab control lane (2).](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/7390_DDX58-Primary-antibodies-ab65588-12.jpg)
DDX58 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to DDX58 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab65588.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 110kDa; DDX58, non-specific band: 60kDa, due to background seen in No ab control lane (2).
Product References
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