Koshland Web Site/Methods
KOSHLAND LAB ANTIBODIES
Antigen Ab Source Western ChIP IF
GFP rabbit Clontech polyclonal 1/>2000 --- 1/4000; 1/1000-2000 in PBS/1%BSA
HA 16B12 (Covance) Mouse (1/1000) --- 1/1000) IP1:150
HA 12CA5 mouse Babco, mouse monoclonal stock -70oC 1/2500; 1/2000 in PBS or TBS, room temp 1-2hrs 8 µl for 1.5ml I.P. o/n at 4 C 1/2000*; 1/100-200 in PBS/3%BSA, room temp at least 2hrs
Mif2 Rabbit Meluh --- 6 µl for 1.5ml I.P. o/n at 4 C ---
Myc Mouse Monoclonal Cohen-Fix 1/5 dilute sera no mild -> in 1%BSA/1XPBST, reuse 10X --- 1/2.5 for spreads
Myc Mouse Monoclonal Santa Cruz 1/2000 in PBS or TBS, room temp 1-2hrs 8 µl for 1.5ml I.P. o/n at 4 C ---
Myc polyclonal Santa Cruz --- 6 µl for 1.5ml I.P. o/n at 4 C ---
Myc Mouse monoclonal Oncogene --- --- 1/500 in PBS/BSA buffer
PAP Sigma 1/2500 --- ---
Pds1 Rabbit Yamamoto --- --- ---
Smc1 Rabbit Strunnikov, C181 --- 100 µl serum, prebound to protein A beads for 1.5ml I.P. o/n at 4 C ---
Smt3 Rabbit Meluh 1/10,000 --- ---
Tub2 YOL1 34 Serotec, Rat, monoclonal --- --- 1/200
Tub2 Rabbit 43 Meluh 1/10,000 --- 1/2000
Tub4 Rabbit Kilmartin 1/10,000
2.5ul in 25mL
 --- 1/5000; 1/10,000 in PBS/1%BSA buffer
V5 mouse Invitrogen mouse Ab (1/5000 for westerns) 10ul in 25mL 1%milk/1%BSA PBST; 1/5000 --- note 10' fix of cells or kill Ag (1/500 for spreads)
Zip1 Rabbit polyclonal --- --- --- 1/100 in PBS/3%BSA, room temp at least 2hrs

2d Source Conc. Western IF FISH
Goat anti Rabbit HRP Biorad 1/2500 --- ---
Goat anti Mouse HRP Biorad 1/2500 (1%BSA, 1% milk, PBST) 5ul/12.5ml ---
Goat anti rat - FITC Jackson --- 1/300 ---
Goat anti mouse - FITC Sigma --- 1/200 1/250
Goat anti Rabbit - FITC --- 1/250 ---
Goat anti Rabbit CY3 --- 1/500 (1/250 for spreads*) ---
Pig anti Goat FITC Jackson --- 1/1000 (1/500 for spreads*) 1/250
Mouse anti Digoxygenin --- --- 1/250
Comments:

Conditions:
alphaHA blots - 1/2500 in 1% BSA, 1% milk PBST

for spreads, same as IIF
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