c-Myc antibody [9E10] (ab32)

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Anonymous (click to contact this user) Abreview submitted on 15 April 2009
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
c-Myc antibody [9E10] (ab32)
Sample	Human Tissue sections (tonsil, cells near crypt)
Specification	tonsil, cells near crypt
Fixative:	Formaldehyde
Antigen retrieval step:	None
Permeabilization:	No
Blocking step:	1% BSA plus 5% normal serum in PBS as blocking agent for  1 hour  · Concentration: 5 % · Temperature: RT°C
Primary Antibody
Dilution	1/500
Incubation time:	16 hours  Temperature: 4°C Diluent: PBS, 1% BSA, azide
Secondary Antibody
Name:	Non-Abcam Antibody was used: biotinylated horse anti-mouse IgG Conjugation: Biotin
Dilution:	1/300
Additional Data
Rating	Good
Additional notes	We noticed that one population of cells in the tonsil was negative with no antigen retrieval, but positive with AR in citrate pH6 (20' in a boiling water bath), so we suggest careful testing of your positive and negative control tissues or cells. Image: Human tonsil near crypt 400x
Image	c-Myc antibody [9E10] for IHC-P (Human) Copyright image used under license

Anonymous Abreview submitted on 13 August 2007
Application:	Western blot
c-Myc antibody [9E10] (ab32)
Sample	Human Cell lysate - whole cell (Hek 293 cells)
Loading amount:	30 µg
Specification	Hek 293 cells
Gel Running Conditions:	Reduced Denaturing
Blocking step:	Milk as blocking agent for  2 hours  · Concentration: 5 % · Temperature: 20°C
Primary Antibody
Dilution	1/5000
Incubation time:	1 hour  Temperature: 20°C Diluent: TBST plus 5% milk
Secondary Antibody
Name:	Abcam antibody: Mouse IgG secondary antibody - H&L
Dilution:	1/5000
Detection
Detection method:	ECL
Exposure:	15 minutes and 0 seconds
Bands:	Specific: 64 kDa
Additional Data
Rating	Excellent

Anonymous (click to contact this user) Abreview submitted on 15 February 2007
Application:	Immunocytochemistry/ Immunofluorescence
c-Myc antibody [9E10] (ab32)
Sample	Human Cell (transfected HEK cells)
Specification	transfected HEK cells
Fixative:	Formaldehyde
Blocking step:	Milk as blocking agent for  20 mins  · Concentration: 3%
Primary Antibody
Dilution	1/250
Incubation time:	45 minutes
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa 488 donkey anti-mouse IgG1 Conjugation: Alexa Fluor® 488
Dilution:	1/500
Additional Data
Rating	Good

review by Gloria Zimmerschied submitted on 17 December 2004
Application:	Western blot
c-Myc antibody [9E10] (ab32)
Additional Data
Notes	1) testing for presence of myc tagged proteins in transfected cells 2) recommended dilutions 1:1000 3) not used on tissue; cell lines only; HCT116 & NIH3T3 4) detection by chemiluminescent secondary 5) we were quite happy with the results 6) no special conditions,diluted stock concentration of 1 mg/ml 1:1000,diluted stock concentration of 1 mg/ml 1:1000
Dilution notes	diluted stock concentration of 1 mg/ml 1:1000

review by Jennifer Edwards submitted on 29 November 2004
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
c-Myc antibody [9E10] (ab32)
Additional Data
Notes	The MYC antibody works nicely! IHC was done on tumor and normal tissues. The staining was intense and we saw very little background. We highly recommend this antibody.
Dilution notes	As suggested by abcam.

review by Sarah Roby submitted on 5 September 2003
Application:	Immunoprecipitation
c-Myc antibody [9E10] (ab32)
Additional Data
Notes	Item used: anit-myc antibody (ab32-100). Used in immunoprecipitations at 1:200. Yes, the anitbody worked very well under normal conditions. I have been using this antibody for several months with no problems, it works very well. Sarah Roby
Dilution notes	IP: 1:200 dilution

review by Claudia Leichtlein submitted on 7 May 2003
Application:	Immunohistochemistry (Frozen sections)
c-Myc antibody [9E10] (ab32)
Additional Data
Notes	I use Abcam c-myc antibody (ab32) for immunohistochemistry with perfused brain sections and in vitro analysis on cell cultures. I have found this antibody to be reliable, long lasting and low background. It is a useful diagnostic tool for my research as it recognises a tagged human protein delivered by viral packaging and does not cross react with any endogenous protein. Good on you Abcam for a superior product.
Dilution notes	1: 1000 (brain sections) 1: 2000 (cell culture)

review by Pietro Pichierri submitted on 1 July 2002
Application:	Immunoprecipitation
c-Myc antibody [9E10] (ab32)
Additional Data
Notes	I have used this antibody in western blot and in immunoprecipitation to detect cMyc-tagged FancG protein For western blot I used the antibody at 1:1000 dilution in TBST+5%NFDM Overnight at 4°C. However, in my hands, this antibody gave a non specific band at approx. 65KDa. The aspecif band is eliminated in immunoprecipitation. In fact, using 3micrograms of antibody for 500micrograms of whole cell lysate, I obtained a clear signal representing the cMyc-tagged protein. I suggest, everytime the tagged protein has a MW near 65KDa, to immunoprecipitate in order to avoid the nonspecific band.
Dilution notes	Western blot: 1:1000 IP: 3micrograms/reaction (up to 0.8micrograms of proteins) .

review by Pietro Pichierri submitted on 11 July 2001
Application:	Western blot
c-Myc antibody [9E10] (ab32)
Additional Data
Notes	This antibody works well either in Western Blotting and Immunofluorescence. I have successfully detected a cMyc-fusion protein transiently expressed in Cos-7 cells using also half of the suggested concentrations.

Anonymous (click to contact this user) Abreview submitted on 19 April 2001
Application:	Western blot
c-Myc antibody [9E10] (ab32)
Additional Data
Notes	The antibody works well for both immunostaining, including double staining, no cross reactions. It works well for western blots, too.