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Fixes&Dilutions

Fixation and Staining Parameters

Antigen

Fixation

Fixation time

Antibody

Dilution

Ref

microtubules

100% MeOH

5 min @ RT

DM1a

1:500

actin

4% Formaldehyde*

8 min @ RT

C4

1:500

1

POD-1

 

Formaldehyde or MeOH*

8 min @ RT

11-1

11-9

1:1000

1:1000

1

PAR-1

100% MeOH

15 min @ -20 deg

PAR-1

1:5

2

PAR-3

100% MeOH

15 min @ RT

PAR-3

1:20

3

P-granules

100% MeOH**

10 min @ 4 deg

K76

1:1 or 1:2

4

* post-fix: 5 min in 100% MeOH
** post-fix: 10 min in 4 deg Acetone and air dry, do not use any detergents

 

 

References

  1. Aroian, RV et al. (1997) EMBO 16:1541-1549
  2. Guo, S and Kemphues, KJ (1995) Cell 81:611-620
  3. Etemad-Moghadam, B et al (1995) Cell 83:743-752
  4. Strome, S and Wood, WB (1983) Cell 35:15-25

 

compiled by Chad Rappleye

Aroian Lab Protocols