Product datasheet Anti-DMD Antibody Catalog Number: PB9276

BOSTER BIOLOGICAL TECHNOLOGY Special NO.1, International Enterprise Center, 2nd Guanshan Road, Wuhan, China

Web: www.boster.com.cn Phone: +86 27 67845390 Fax: +86 27 67845390 Email: [email protected]

Basic Information Product Name Anti-DMD Antibody Gene Name DMD Source Rabbit IgG Species Reactivity human,mouse,rat Tested Application WB,IHC-P,IHC-F,ICC,FCM Contents 500ug/ml antibody with PBS ，0.02% NaN3 , 1mg BSA and 50% glycerol. Immunogen E.coli-derived human recombinant (Position: H3076-D3404). Human Dystrophin shares 100% amino acid (aa) sequence identity with mouse Dystrophin. Purification Immunogen affinity purified. Observed MW 427KD Dilution Ratios Western blot: 1:500-2000 Immunohistochemistry(Paraffin-embedded Section): 1:50-400 Immunohistochemistry(Frozen Section): 1:50-400 Immunocytochemistry: 1:50-400 Flow Cytometry: 1-3μg/1x106 cells (Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user.

Storage 12 months from date of receipt，-20℃ as supplied.6 months 2 to 8℃ after reconstitution. Avoid repeated freezing and thawing

Background Information Dystrophin, also known as DMD, is a rod-shaped cytoplasmic protein, and a vital part of a protein complex that connects the of a muscle fiber to the surrounding extracellular matrix through the cell membrane. It is mapped to Xp21.2-p21.1. This complex is variously known as the or thedystrophin-associated protein complex. Many muscle , such as α-, , , , , and , colocalize with dystrophin at the costamere. Dystrophin is a protein located between the and the outermost layer of in the muscle fiber (myofiber). It is a cohesive protein, linking filaments to another support protein that resides on the inside surface of each muscle fiber’s plasma membrane (sarcolemma).

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FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 1 Product datasheet Anti-DMD Antibody Catalog Number: PB9276

BOSTER BIOLOGICAL TECHNOLOGY Special NO.1, International Enterprise Center, 2nd Guanshan Road, Wuhan, China

Web: www.boster.com.cn Phone: +86 27 67845390 Fax: +86 27 67845390 Email: [email protected]

Selected Validation Data

Figure 1. Western blot Figure 2. IHC analysis of analysis of Dystrophin Dystrophin using anti- using anti-Dystrophin Dystrophin antibody antibody (PB9276). (PB9276).Dystrophin was Electrophoresis was detected in paraffin- performed on a 5-20% embedded section of SDS-PAGE gel at 70V Mouse Brain Tissue. Heat (Stacking gel) / 90V mediated antigen (Resolving gel) for 2-3 retrieval was performed hours. The sample well of in citrate buffer (pH6, each lane was loaded epitope retrieval solution) with 50ug of sample for 20 mins. The tissue under reducing section was blocked with conditions. Lane 1: 10% goat serum. The human HEK293 whole tissue section was then cell lysates, Lane 2: incubated with 1μg/ml human K562 whole cell rabbit anti-Dystrophin lysates, Lane 3: mouse Antibody (PB9276) HEPA1-6 whole cell overnight at 4°C. lysates, After Biotinylated goat anti- Electrophoresis, proteins rabbit IgG was used as were transferred to a secondary antibody and Nitrocellulose membrane incubated for 30 minutes at 150mA for 50-90 at 37°C. The tissue minutes. Blocked the section was developed membrane with 5% Non- using Strepavidin-Biotin- fat Milk/ TBS for 1.5 hour Complex (SABC)(Catalog at RT. The membrane was # SA1022) with DAB as incubated with rabbit anti- the chromogen. Dystrophin antigen affinity purified polyclonal antibody (Catalog # PB9276) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti- rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Dystrophin

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 2 Product datasheet Anti-DMD Antibody Catalog Number: PB9276

BOSTER BIOLOGICAL TECHNOLOGY Special NO.1, International Enterprise Center, 2nd Guanshan Road, Wuhan, China

Web: www.boster.com.cn Phone: +86 27 67845390 Fax: +86 27 67845390 Email: [email protected]

at approximately 427KD. The expected band size for Dystrophin is at 427KD.

Figure 3. IHC analysis of Figure 4. IHC analysis of Dystrophin using anti- Dystrophin using anti- Dystrophin antibody Dystrophin antibody (PB9276).Dystrophin was (PB9276).Dystrophin was detected in paraffin- detected in paraffin- embedded section of Rat embedded section of Cardiac . Human Lung Cancer Heat mediated antigen Tissue. Heat mediated retrieval was performed antigen retrieval was in citrate buffer (pH6, performed in citrate epitope retrieval solution) buffer (pH6, epitope for 20 mins. The tissue retrieval solution) for 20 section was blocked with mins. The tissue section 10% goat serum. The was blocked with 10% tissue section was then goat serum. The tissue incubated with 1μg/ml section was then rabbit anti-Dystrophin incubated with 1μg/ml Antibody (PB9276) rabbit anti-Dystrophin overnight at 4°C. Antibody (PB9276) Biotinylated goat anti- overnight at 4°C. rabbit IgG was used as Biotinylated goat anti- secondary antibody and rabbit IgG was used as incubated for 30 minutes secondary antibody and at 37°C. The tissue incubated for 30 minutes section was developed at 37°C. The tissue using Strepavidin-Biotin- section was developed Complex (SABC)(Catalog using Strepavidin-Biotin- # SA1022) with DAB as Complex (SABC)(Catalog the chromogen. # SA1022) with DAB as the chromogen.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 3

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