Application
Immunoprecipitation
Sample
Human Cell lysate - other (Human serum)
Total protein in input
500 µg
Immuno-precipitation step
Other - Protein A and G Dynabeads and Protein A Agarose
Specification
Human serum
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 50mM Ammounium acetate
Dilution
1/4
Western Blot antibody
Western Blot antibody
None used
Additional data
Additional Notes
We trialled this antibody for IP but unfortunately were not able to achieve immunodepletion of our 8.6 kDa target (C4a anaphylotoxin). We attempted this via a number of approaches, namely:
- protein G Dynabeads, indirect and direct
- protein A Dynabeads, indirect
- protein A agarose beads, indirect and direct.
In every case we followed the manufacturer's instructions for the Dynabeads or the agarose resin, but, after assessment on our SELDI-TOF mass spectrometery, no depletion of this protein was seen.
We have uploaded an image of the SELDI spectra of serum pre- and post-depletion by IP using 2ug of antibody (indirectly with Dynabeads G in this case), and you can see that there is no decrease in the intensity of the peak at 8.6 kDa.
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. Shea Hamilton
Verified customer
提交于 Sep 30 2016