Recombinant Anti-delta Sarcoglycan antibody [EPR8706] (ab137101)

Fluorescence multiplex immunohistochemical analysis of the human cardiac muscle (Formalin/PFA-fixed paraffin-embedded sections).

Panel A: merged staining of anti-delta Sarcoglycan (ab137101, red; Opal™690), anti-Titin (ab307446, green; Opal™520) and anti-Natriuretic peptides A (ab209232, magenta; Opal™570) on human cardiac muscle. Panel B: anti-Titin displayed nucleus and cytoplasm expression. Panel C: anti-Natriuretic peptides A displayed granular cytoplasmic expression. Panel D: anti-delta Sarcoglycan displayed membrane expression. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

The section was incubated in three rounds of staining: in the order of ab137101 at 1/1000 (1.043 μg/ml) dilution, ab307446 at 1/500 (0.95 μg/ml) dilution, and ab209232 at 1/3000 (0.241 μg/ml) dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI (blue) was used as a nuclear counter stain.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Fluorescence multiplex immunohistochemical analysis of the human skeletal muscle (Formalin/PFA-fixed paraffin-embedded sections).

Panel A: merged staining of anti-delta Sarcoglycan (ab137101, green; Opal™690), anti-Fast Myosin Skeletal Heavy chain + MYH4 (ab221149, magenta; Opal™520) and anti-Slow Skeletal Myosin Heavy chain (ab234431, red; Opal™570) on human skeletal muscle. Panel B: anti-Fast Myosin Skeletal Heavy chain + MYH4 stained on fast type fibers. Panel C: anti-Slow Skeletal Myosin Heavy chain stained on slow type fibers. Panel D: anti-delta Sarcoglycan stained on membrane of skeletal muscle. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

The section was incubated in three rounds of staining: in the order of ab137101 at 1/1000 (1.043 μg/ml), ab221149 at 1/1000 (0.505 μg/ml, and ab234431 at 1/4000 (0.255 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI (blue) was used as a nuclear counter stain.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Fluorescence multiplex immunohistochemical analysis of the human skeletal muscle (Formalin/PFA-fixed paraffin-embedded sections).

Panel A: merged staining of anti-delta Sarcoglycan (ab137101, red; Opal™690), anti-MYH2 (ab264036, cyan; Opal™520) and anti-MYH6 + Slow Skeletal Myosin Heavy chain (ab250868, gray; Opal™570) on human skeletal muscle. Panel B: anti-MYH2 stained on the fast twitch type 2A fibers. Panel C: anti-MYH6 + Slow Skeletal Myosin Heavy chain stained on skeletal muscle fiber subtypes. Panel D: anti-delta Sarcoglycan stained on membrane of skeletal muscle. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

The section was incubated in three rounds of staining: in the order of ab137101 at 1/1000 dilution (1.043 μg/ml), ab264036 at 1/1000 dilution (1.116 μg/ml) and ab250868 1/5000 dilution (0.209 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI (blue) was used as a nuclear counter stain.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Human heart tissue labelling delta Sarcoglycan with ab137101 at 1/100 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Purified ab137101 at 1/20 dilution (1µg) immunoprecipitating delta Sarcoglycan in Human skeletal muscle lysate.
Lane 1 (input): Human skeletal muscle lysate 10µg
Lane 2 (+): ab137101 + Human skeletal muscle lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab137101 in Human skeletal muscle lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 35 kDa