Hi Nagarajan,

Beige ( bg) mutation in the mice leads to complete impairment in NK cell function .

You may mind procedure for identification of Beige mice progeny by going through the procedures described in the following papers published in

a) Jr. of Immunology Vol 123(5),2174-2181 , 1979

b) Am. J. Pathol.Vol 82(3 ), 573-588 ,March,1976

You can discuss about this with Dr.Rath/Dr.Bal abd they may help you in setting up an immunoassay for bg mice identification.

When you have mice with beige gene mutation, you can phenotypically identify the mice on the basis of their coat color.

beige color is a lighter / diluted version of black color .

You can google beige color photographs on the net which will give you idea how mice with beige phenotype looks like .

Regards,

Rajesh Anand

Thanks a lot for sending the Reference .I will check accordingly

Hi Dr.Nagarajan,

You can use this procedure to identify homozygous beige ( bg/bg ) mice genotype population from heterozygote beige ( +/bg ) and the non-beige ( +/+ ) genotype mice .

Identification ofthe bg/bg genotype : Beige mice are most readily identified by staining blood films and detecting the presence of giant sudanophilic granules in 100% of their blood neutrophils and by coat colour dilution, in non-white animals. Smears of blood were prepared from all animals by touching a clean glass slide to the incised tail vein of each animal then spreading the drop using a second glass slide. Slides were air dried then fixed in paraformaldehyde vapour for 10 min and washed in running tap water for 10 min. Next, slides were immersed in a filtered staining solution made by mixing 60 ml of a 0-3% (w/v) solution of Sudan black (Sigma) in 95% ethanol with 40 ml of a buffered solution made of 53% (w/v) crystalline phenol in 95% ethanol mixed at 1:3 with a 0-3% w/v aqueous solution of Na2HP04.12H20 for 60 min. The slides were then dipped 5 times, for 10 sec each dip, in 50% ethanol, washed for 5 min in running tap water and blown dry. The slides were counterstained with Gill's haematoxylin for 3 min, blown dry, and examined using a light microscope at 400 or 1000 oil magnification. At least 5

neutrophils were examined from each animal and scored for granule size. Mice with blood smears displaying giant granules were designated bg/bg, and this coincided with coat colour dilution in all of the non-white mice. Mice with blood films displaying neutrophil granules of normal size were designated non-beige, and may have been +/bg or + / +. The non-beige animals have been designated + /bg(?).

hanks ,I planned to check the blood smear along with FACS to know the NK1.1 population of screened Mice positive for Prkdc gene to know the beige .

Nagarajan