Secondary antibodies

Secondary antibodies kdorfman Tue, 10/02/2018 - 19:19

Goat anti-mouse IgM TRITC

Goat anti-mouse IgM TRITC kdorfman Tue, 10/02/2018 - 19:26

Fisher OB102102 (Southern BioTech 1021-02)

Goat Anti-Mouse IgM (micron chain specific)-FITC

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 - 20:37

Invitrogen A-11001

Ex: 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 - 19:23

Anti-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 FITC

Goat anti-rabbit FITC kdorfman Tue, 10/02/2018 - 20:42

Goat anti-rabbit IgG FITC

Sigma F0382

Dilute 1:80

Goat anti-rabbit red

Goat anti-rabbit red kdorfman Tue, 10/02/2018 - 19:25

Jackson 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 - 19:24

Sigma 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 - 19:31

From 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)