ECT2 antibody (ab4863)

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review by JIHYE HAN submitted on 29 January 2010
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
ECT2 antibody (ab4863)
Sample	Human Tissue sections (colon)
Specification	colon
Fixative:	Formaldehyde
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: 0.01M Tris Buffer (pH9.0)/ 0.01M citrate Buffer (pH6.0)
Permeabilization:	No
Blocking step:	BSA as blocking agent for  1 hour  · Concentration: 4 % · Temperature: 25°C
Primary Antibody
Dilution	1/20
Incubation time:	1 hour  Temperature: 25°C Diluent: PBST
Secondary Antibody
Name:	Non-Abcam Antibody was used: Anti-Goat polymer kit (HRP-conjugated ?-Goat Ig ) Conjugation: HRP
Additional Data
Rating	Inconclusive
Additional notes	Primary Antibody diluent 1:100~1:20 strong background, no staining at Nuclear(negative)
Image	ECT2 antibody for IHC-P (Human) Copyright image used under license
Abcam response	Customer tried different antigen retrievals with pressure cooker for 30min. ECT2 should have nuclear localization in interphase, spreads throughout the cytoplasm in prometaphase, and is condensed in the midbody during cytokinesis.

Anonymous (click to contact this user) Abreview submitted on 23 July 2009
Application:	Immunocytochemistry/ Immunofluorescence
ECT2 antibody (ab4863)
Sample	Human Cell (HeLa)
Specification	HeLa
Fixative:	Methanol
Permeabilization:	No
Blocking step:	BSA as blocking agent for  30 mins  · Concentration: 2 % · Temperature: 27°C
Primary Antibody
Dilution	1/50
Incubation time:	1 hour  Temperature: 27°C
Secondary Antibody
Name:	Non-Abcam Antibody was used: Donkey anti-Goat Conjugation: Alexa Fluor® 594
Dilution:	1/500
Additional Data
Rating	Poor
Additional notes	We tried various conditions and also different fixations (MeOH, Formaldehyde, Paraformaldehyde, TCA) but could not get any signal from this antibody. It's not on midzones/midbody/furrow during cytokinesis as in published papers. Also we didn't see it in interphase nucleus as the picture shown in the website.
Image	ECT2 antibody for ICC/IF (Human) Copyright image used under license
Abcam response	This negative Abreview is currently being investigated by Abcam Technical Support. Please check here for an update response from Abcam.

review by NASLIN RASHEED submitted on 29 April 2005
Application:	Immunofluorescence
ECT2 antibody (ab4863)
Additional Data
Notes	I used anti-Ect2 goat polyclonal for immuno fluourescence staining. It was used at a dilution of 1:500 in 1%BSA-PBS solution. The incubation was carried out for 2 hours at R.T. I was able to observe staining of Ect2 in my cells, consistent with the staining obtained using anti-Ect2 rabbit polyclonal.
Dilution notes	1:500 in 1%BSA/PBS

review by Jelissa Cheng submitted on 13 October 2004
Application:	Western blot
ECT2 antibody (ab4863)
Additional Data
Notes	Antibody works for western analysis (using human U87-MG glioblastoma) but signal is a little weak for the 100kDa band. Small 30+ kDa band observed as well. Relatively little unspecific signal.
Dilution notes	1:200 for primary antibody, 1:500 for secondary antibody