On January 5, 2017Seandean Harwood, Aarhus University wrote:
5
M2 anti-FLAG tag (murine IgG1)
Product # F3165 from Sigma-Aldrich
This is the standard antibody which I have used for a variety of applications in detecting several recombinant FLAG-tagged proteins. Very reliable.
ELISA: used at 0.5 ug/mL in 2% BSA PBS, followed by detection with a GAM-HRP
Western blot: used at 0.5 ug/mL in 2% milk powder – PBS, then detected with GAM-HRP
FACS: used at 5 ug/mL in 1% FBS PBS, then detected with GAM-PE
Western blot
Immunostaining
Not Rated
IP
Not Rated
ELISA
Flow cytometry
Luminex
Not Rated
On March 25, 2015Morten Callesen, Biomed AU wrote:
3
Western Blot detection of transfected HEK293T transient expressing FLAG tagget Cas9 protein. Only Faint unspecific signal on 30µg/lane (1:5000 HRP pRb)
Dilution 1:2000
Western blot
Immunostaining
Not Rated
IP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On December 23, 2014Julie Torvund-Jensen, Aarhus University wrote:
4
This antibody works fine for the detection of a C-terminal FLAG-tagged protein in rat primary oligodendrocytes.
I have used this antibody against a FLAGtag in the c-terminal of a receptor protein, with some success. The antibody needs to be fresh, as the stock in the fridge did not work. A fresh stock from -20C worked great, with comparable signal strength to an antibody against the actual receptor protein. The M2 ab is Calcium independent and bind FLAG tag anywhere in the protein.
Dilut 1:1000.
Western blot
Immunostaining
Not Rated
IP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On December 2, 2013Sune Skeldal wrote:
0
This antibody dimerises/polymerises exogenous FLAG-tagged sFasL in cell cultures, which dimerises/polymerises FAS in jurkat cells. I used 1 ug/mL in cell culture medium for 5 hours at 37 C.
On January 27, 2013Simon Glerup wrote:
4
Works for the detection of FLAG-tagged proteins expressed in HEK293 cells by immunofluorescense.
Dilution 1:500
On January 5, 2017 Seandean Harwood, Aarhus University wrote:
M2 anti-FLAG tag (murine IgG1)
Product # F3165 from Sigma-Aldrich
This is the standard antibody which I have used for a variety of applications in detecting several recombinant FLAG-tagged proteins. Very reliable.
ELISA: used at 0.5 ug/mL in 2% BSA PBS, followed by detection with a GAM-HRP
Western blot: used at 0.5 ug/mL in 2% milk powder – PBS, then detected with GAM-HRP
FACS: used at 5 ug/mL in 1% FBS PBS, then detected with GAM-PE
On March 25, 2015 Morten Callesen, Biomed AU wrote:
Western Blot detection of transfected HEK293T transient expressing FLAG tagget Cas9 protein. Only Faint unspecific signal on 30µg/lane (1:5000 HRP pRb)
Dilution 1:2000
On December 23, 2014 Julie Torvund-Jensen, Aarhus University wrote:
This antibody works fine for the detection of a C-terminal FLAG-tagged protein in rat primary oligodendrocytes.
Dilution 1:100
On December 2, 2013 Mads Kjolby wrote:
I have used this antibody against a FLAGtag in the c-terminal of a receptor protein, with some success. The antibody needs to be fresh, as the stock in the fridge did not work. A fresh stock from -20C worked great, with comparable signal strength to an antibody against the actual receptor protein. The M2 ab is Calcium independent and bind FLAG tag anywhere in the protein.
Dilut 1:1000.
On December 2, 2013 Sune Skeldal wrote:
This antibody dimerises/polymerises exogenous FLAG-tagged sFasL in cell cultures, which dimerises/polymerises FAS in jurkat cells. I used 1 ug/mL in cell culture medium for 5 hours at 37 C.
On January 27, 2013 Simon Glerup wrote:
Works for the detection of FLAG-tagged proteins expressed in HEK293 cells by immunofluorescense.
Dilution 1:500
On January 12, 2013 Simon Glerup wrote:
Works OK for IP