Secondary antibodies
Secondary antibodies kdorfman Tue, 10/02/2018 - 19:19Goat anti-mouse IgM TRITC
Goat anti-mouse IgM TRITC kdorfman Tue, 10/02/2018 - 19:26Fisher OB102102 (Southern BioTech 1021-02)
Goat Anti-Mouse IgM (micron chain specific)-TRITC
1 mg/mL
1:100 - 1:400
1 µL aliquots. Add 100 - 400 µL 1% BSA
Use 50 µL per coverslip
Goat anti-mouse 488
Goat anti-mouse 488 kdorfman Tue, 10/02/2018 - 20:37Ex: 488 nm
Em: 499 nm
Stock(as purchased) = 2 mg/mL
Stock bottle has been diluted with glycerol: ~800 uL of 1 mg/mL
Working concentration = 1 ug/mL
Aliquots are 1 uL of 1 mg/mL
Add 611 uL to make enough for 6 slides, or up to 1 mL
Freezer 266A
Goat anti-mouse TRITC
Goat anti-mouse TRITC kdorfman Tue, 10/02/2018 - 19:23Anti-Mouse IgG (whole molecule) TRITC conjugate
Sigma T 5393
26 mg protein/mL
minimum working dilution = 1:64
1 µL aliquots. Add up to 100 µL 1% BSA
Alexa Fluor 568 (A11004) (red) 2mg/mL
working concentration ~10µg/mL
dilute 1:200
aliquots are 110 µL 20µg/mL
add
110 µL 2% BSA in PBS
to make 220 µL 10µg/mL
Use 50 µL per coverslip (there is enough for 4 per tube)
Goat anti-rabbit 546
Goat anti-rabbit 546 kdorfman Tue, 10/02/2018 - 20:39Stock concentration = 2 mg/mL
Working concentration = 4 ug/mL
Em = 575 nm
Ex = 560 nm
ALL OUT as of Spring 2018
Goat anti-rabbit FITC
Goat anti-rabbit FITC kdorfman Tue, 10/02/2018 - 20:42Goat anti-rabbit red
Goat anti-rabbit red kdorfman Tue, 10/02/2018 - 19:25Jackson Immuno 111-165-003
Cy3 Ex - 550 nm, em= 570 nm ("Red")
tubes : 2 µL. Add 398 µL PBS-Tw-Az to make 1:200
Use 50 µL per coverslip (there is enough for 4 per tube)
Goat anti-rat FITC
Goat anti-rat FITC kdorfman Tue, 10/02/2018 - 19:24Sigma F6258
Each tube has 3 µL antibody
Add
192 µL PBS-Tw-Az
195 µL 2% BSA
=390 µL, enough for 7 reactions (50 µL/coverslip 37C 30min)
Goat anti-rat TRITC
Goat anti-rat TRITC kdorfman Tue, 10/02/2018 - 19:31From Sigma AP136R EMB Millipore
ab peak = 550nm; emission peak = 570 nm
Bottle contains 2 mg. Add 1 mL H2O + 1 mL glycerol to make 1 mg/mL
aliquot 2 µL or 5 µL antibody, freeze
add 198 µL or 495 µL PBS-Tw-BSA
use 50 µL per coverslip, diluted to 1/100 (Try 37C 30 min)