PA5-39018

antibody from Invitrogen Antibodies

Targeting: SGCA A2, adhalin, ADL, DMDA2, LGMD2D, SCARMD1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-39018

Provider

Invitrogen Antibodies

Product name

Anti-Alpha Sarcoglycan Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of alpha Sarcoglycan/SGCA in extracts from Jurkat cells, HepG2 cells and HeLa cells using an Alpha Sarcoglycan/SGCA polyclonal antibody (Product # PA5-39018).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Alpha Sarcoglycan was achieved by transfecting MCF7 with Alpha Sarcoglycan specific siRNAs (Silencer® select Product # S12764, S12763). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the Alpha Sarcoglycan knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Alpha Sarcoglycan Polyclonal Antibody (Product # PA5-39018, 1:2000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). Densitometric analysis of this western blot is shown in the histogram (Fig. b). The decrease in signal upon siRNA mediated knockdown confirms that the antibody is specific to Alpha Sarcoglycan.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Alpha Sarcoglycan Polyclonal Antibody(Product # PA5-39018) and 43-55 kDa bands corresponding to Alpha Sarcoglycan were observed across cell lines and tissues tested. An uncharacterized band (*) was observed around 10 kDa. Membrane enriched extracts (30ug lysate) of A549 (Lane 1), HEL 92.1.7 (Lane 2), BeWo (Lane 3) and tissue extracts of Rat Skeletal Muscle (Lane 4), Mouse Heart (Lane 5), Rat Heart (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Alpha Sarcoglycan was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Alpha Sarcoglycan Polyclonal Antibody (Product # PA5-39018) at 1:100 dilution in 0.1% BSA, incubated at 4-degree Celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Plasma membrane and cytoplasm localization. Panel e represents control cells with no primary antibody to assess the background. The images were captured at 60X magnification.