Hmmm.

Not sure at all.

ab32077/ab52894 have a similar pattern of positivity.

Comments, please?

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?

Hey…comments, please.

A very similar pattern of positivity is seen when detecting using ab32077 ( further dilution is reqd)

This reagent should be further diluted.

Most intense positivity ( membraneous) is seen in germinal cell layer of developing epithelium and also in hair papillae

Should the positivity with this phosphorylated antibody be ( seemingly) identical to that seen when probing the same cell line with ab32077/52894?

Same comments apply here as with ab32077

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.

Comments added to ab32077 apply here.

Help/comments would be much appreciated.

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

🙂

The megakaryocytes are positive: I do not know if this is specific.  Using anti EGFr ab32077/ab52894 these cells are negative, under same detection conditions.