Not sure at all.
ab32077/ab52894 have a similar pattern of positivity.
Positivity seen with top two images( ? total EGFr?) is similar: a subset of tubular and also interstitial positivity.
However, using ab40815 ( phosphorylated- lowest image) there is clear membraneous positivity in transitional urothelium ( central feature) and, I assume, collecting ducts?
Histonet is BIG so….look at images/add images and please help in gaining a better understanding
Antibody should be further diluted ( used at 1/400)
Cells were both scraped or trypsinised ( in case enzyme altered signal), spun down. Resuspended in 10 % Formalin O/N at RT.
Spun down, rinsed/resuspended in TBS.
After another spin, cell pellet was resuspended in HMP Agarose at 50C and then immediately spun down at 4K for 5 mins.
Agarose gel containing pellet was processed to Pwax conventionally.
Is this positivity specific?
It appears to be identical to that seen using ab40815 and also ab52894
Sure, one sees membrane positivity.
I am not familiar with EGFr positivity/distribution within any tissues under these conditions.
However, HPA shows an identical positivity under similar conditions. Abcam’s ab52894 shows a very similar distribution pattern.
However, Abcam’s ab40815 (anti pEGFr) is negative.
Comments always welcomed