Anti-Dystrophin 抗体

• IHC

Lab

Immunohistochemistry - Anti-Dystrophin antibody (AB15277)

Immunohistochemical analysis of formalin fixed paraffin embedded human heart labelling Dystrophin with ab15277 at a dilution of 1/800. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32 mins.

ab15277 Anti-Dystrophin antibody was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dystrophin antibody (AB15277)

Dystrophin quantification in a population of myofibres identified in entire muscle sections performing the double labelling anti-dystrophin ab15277 (red; 1/200 dilution) and anti-spectrin (green; 1/20 dilution).

All the labellings were performed at RT. Human Muscle sections were incubated with the primary antibody combination (anti-dystrophin ab15277 and anti-spectrin) for 1 hour. After three washes with PBS sections were incubated with Alexa Fluor 488 conjugated anti-mouse IgG (1 : 100, Thermo Fisher Scientific, Hemel Hempstead, UK) and anti-rabbit biotinylated IgG (1 : 200; GE Healthcare, Amersham Pl, UK) for 30 minutes. PBS washes were performed and sections were incubated with Alexa Fluor 594 streptavidin conjugate (1 : 1000, Thermo Fisher Scientific, Hemel Hempstead, UK).

Representative images of entire muscle sections stained and acquired by the Axio Scan slide scanner and processed with Definens algorithm derived from a control (a) and from a DMD patient (b).

DMD : Duchenne Muscular Dystrophy.

Sardone V. PLoS One. 2018 Mar 26;13(3):e0194540. doi: 10.1371/journal.pone.0194540. eCollection 2018. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dystrophin antibody (AB15277)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human skeletal muscle labeling Dystrophin with ab15277 at 1/100 dilution.

• IHC-Fr

Project24781921****

Immunohistochemistry (Frozen sections) - Anti-Dystrophin antibody (AB15277)

Immunofluorescence staining of dystrophin in W9, W987, and ESC. Myosin heavy chain (MHC) identified mouse muscle cells after differentiation. DAPI was used to stain nuclei.

Seventy-two hours before engraftment, 8 week-old mdx/SCID mice received 14 Gy of irradiation localized to the hind limb muscles. On the day of engraftment, SM/C-2.6-positive myogenic cells were purified by fluorescence-activated cell sorting (FACS), using a BD Aria II FACS machine and the same labeling protocol as described above for FC analysis, resuspended in 30 μl of phosphate buffered saline (PBS), loaded into an insulin syringe (BD), and injected into the left tibialis anterior (TA) muscle of anesthetized mice. 7.5×10⁵ differentiated and sorted W987 cells were injected. Control mice were injected with PBS alone. Three weeks following engraftment, TA muscles were harvested, fixed in 0.5% paraformaldehyde for 4 hours, dehydrated in 20% sucrose overnight and frozen in optimal cutting temperature (OCT) using liquid nitrogen cooled methyl-butane. Tissue blocks imbedded in OCT were cryosectioned and processed for immunocytochemical analysis using rabbit anti-dystrophin. Secondary antibodies used were donkey anti-rabbit conjugated to Alexafluor 594 and donkey anti-rat conjugated to Alexafluor 488 (Life Technologies). Nuclei were visualized using NucBlue Fixed Cell Stain (Life Technologies).

Gene-corrected mdx iPSC W987, non-gene-corrected unexcised mdx iPSC W9 and wild-type ESC controls.

Zhao C. et al PLoS One. 2014 Apr 29;9(4):e96279. doi: 10.1371/journal.pone.0096279. eCollection 2014.

• IHC-Fr

AbReview

Immunohistochemistry (Frozen sections) - Anti-Dystrophin antibody (AB15277)

Muscle stem cells (from normal mouse) were injected into the gastric muscle of an MDX mouse. Dystrophin staining : primary antibody ab15277 and secondary antibody is donkey anti-rabbit Alexa 594.

This image was kindly supplied as part of the review submitted by Jessica Tebbets.