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Table 1 List of antibodies and conditions in which they were used

From: MuscleJ: a high-content analysis method to study skeletal muscle with a new Fiji tool

 

Primary antibody

Secondary antibody

Fiber/nuclei (from 2 labs)

Laminin

Laminin α-2 (4H8-2) Santa Cruz #59854 (1/250) ON 4 °C

Donkey anti-rat IgG Alexa Fluor 488 ThermoFisher #A-21208 (1/500) 45 min RT

Hoechst

 

Hoechst 33342 ThermoFisher #H3570 (1/1500) 45 min RT

Laminin

Anti-laminin Ab (produced in Rabbit) L9393 thermoscientific (1/100) ON 4 °C

Pierce donkey anti-rabbit IgG (H+L) cross adsorbed secondary Ab DyLight 650 (prod#Sa5-1041) (1/200) 1 h RT

Hoechst

 

Hoechst 33342 ThermoFisher #H3570 (1/1000) 1 h RT

Satellite cell

Laminin

Anti-laminin Ab (produced in rabbit) L9393 thermoscientific (1/100) ON 4 °C

Pierce donkey anti-rabbit IgG (H+L) cross adsorbed secondary Ab DyLight 650 (prod#Sa5-1041) (1/200) 1 h RT

Pax7

Monoclonal mouse anti-Pax7(supernatant), Developmental Studies Hybridoma Bank (1/20) ON 4 °C

Biotinylated horse anti-mouse IgG (H + L) (Vector Laboratories, #BA-2000) = amplification (1/200) 1 h RT + Streptavidin-DTAF (Beckman Coulter, #PN IM0307) (1/1000) 1 h RT

Vessels

Laminin

Anti-laminin Ab (produced in rabbit) L9393 thermoscientific (1/100) ON 4 °C

Pierce donkey anti-rabbit IgG (H + L) cross adsorbed secondary Ab DyLight 488 (prod#Sa5-10038)

CD31

BD Pharmigen 550,274 purified rat anti-mouse CD31 (1/50) ON 4 °C

Pierce donkey anti-rat IgG (H + L) cross adsorbed secondary Ab DyLight 550 (Prod#Sa5-10027) (1/200) 1 h RT

Fiber type

Type I

BA-D5 (mouse IgG2b) DSHB (1/100) 1 h 37 °C

Anti-mouse IgG2b 647 nm life technology A21242 (1/250) 30 min 37 °C

Type IIA

SC-71 (mouse IgG1) DSHB (1/100) 1 h 37 °C

Anti-mouse IgG1 568 nm life technology A21124 (1/250) 30 min 37 °C

Type IIB

BF-F3 (mouse IgM) DSHB (1/100) 1 h 37 °C

Anti-mouse IgM 488 nm life technology A21042 (1/250) 30 min 37 °C

Laminin

anti-laminin (rabbit) DSHB(1/100) 1 h 37 °C

Anti-rabbit 4+A2:D1905nm abcam ab175651 (1/250) 30 min 37 °C