Novus Biologicals
EGLN3/PHD3 Antibody
NB100-139
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Browse Similar Products: PHD3 Western Blot Antibodies
Goal: The goal was to determine induction of PHD3 in primary murine bone marrow derived macrophages stimulated with a cytokine. Macrophages were left untreated (left lane) or treated with cytokine (right lane) for 24h. After this time, cell lysates were prepared to probe for PHD3 protein. This product was chosen because it has been tested in human cell lines and has been cited over 30 times. Procedure: Cell lysates prepared from murine primary bone marrow derived macrophages were run on a 4-20% gradient gel and transferred onto a 0.2 um nitrocellulose membrane. Membranes were probed with rabbit PHD3 antibody (1:1000) in 5% skim milk at 4°C overnight on a shaker. Blots were washed in TBST 3X and probed with secondary goat anti-rabbit peroxidase (1:1000) for 2 h at ambient temperature followed by washing 3X in TBST. The western blot was developed with Supersignal West Femto Maximum Sensitivity Substrate (Thermo Fisher). Image shows two lanes with PHD3 band seen at ~30 kDa.
Western Blot
Mouse macrophage lysates
1:1000 dilution, overnight, 4 deg
5% skim milk
2h, Room temperature, HRP labelled Ab
None
Supersignal West Femto Maximum Sensitivity Substrate (Thermo Fisher)
PHD3 protein stains at ~30 kD. Some non-specific bands are seen, but overall, detection is good for PHD3.
N/A
Works well for mouse
Some non-specific bands also seen
Can be used successfully in primary mouse cells expressing PHD3.