Application
Immunoprecipitation
Sample
Staphylococcus aureus Cell lysate - whole cell (Primmary Osteoblasts cells)
Total protein in input
200000 cells
Specification
Primmary Osteoblasts cells
Immuno-precipitation step
Other - Dynabeads 280 (invitrogen)
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: PBS 1% w/o mg and Ca
Western Blot antibody
Western Blot antibody
None used
Additional data
Additional Notes
The antibody was tested at first with different concentrations of S. aureus alone and it was really effective. We could recover aprox. 80-90% of the bacterial population. We tested then to recover S. aureus from Primmary osteoblasts after 3h of infection but we could recover only 5% of the total population. After the infection, we lysated the cells which contain S. aueus and we incubated 1h with the antibody and then we added the dynabeads to recover the bacteria-Igs complex and incubate 1h more. All the incubations were at RT with rotation. We think that the unespecific binding was the problem.
Abcam response
We would like to thank the reviewer for submitting this Abreview for product ab20920. As this product had not yet been tested in IP, we were unsure what the result might be and we appreciate the time and effort that the reviewer has put in to testing this product in this application. The customer has received a free primary antibody of their choice for performing these tests. We wish to thank them for participating in our 100% Abreview testing discount program.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Dr. Lorena Tuchscherr
Verified customer
投稿 Feb 23 2012