Western blot analysis of extracts of A-431 and HUVEC cells, using Phospho-EGFR-T669 antibody (AP0025) at 1:1000 dilution. A431 cells were treated by EGF (100ng/mL) for 30 minutes after serum-starvation overnight. HUVEC cells were treated by EGF.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.
Immunoprecipitation analysis of 200ug extracts of A-431 cells, using 3 ug Phospho-EGFR-T669 pAb (AP0025). Western blot was performed from the immunoprecipitate using Phospho-EGFR-T669 pAb (AP0025) at a dilition of 1:1000. A-431 cells were treated by EGF (100 ng/mL) at 37℃ for 30 minutes after serum-starvation overnight.
|Product name||Phospho-EGFR-T669 Rabbit pAb|
|Purification method||Affinity purification|
|Tested applications||WBIHCICCIFIPChIPChIP-seqRIPFCELISAMeDIPNucleotide ArrayDBFACSCoIP|
|Recommended dilution||WB 1:500 - 1:2000|
IP 1:50 - 1:100
|Storage buffer||Store at -20℃. Avoid freeze / thaw cycles.|
Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.
|Application key||Western blotting Immunoprecipitation|
|Positive samples||A-431, HUVEC|
|Cellular location||Cell membrane, Endoplasmic reticulum membrane, Endosome, Endosome membrane, Golgi apparatus membrane, Nucleus membrane, Nucleus, Secreted, Single-pass type I membrane protein|
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