- PA5-52601 - Invitrogen Antibodies SMARCA5 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of SMARCA5 in Rh30 cells transfected with control siRNA, target specific siRNA probe #1 and #2, using a SMARCA5 Polyclonal Antibody (Product # PA5-52601). Remaining relative intensity is presented. Loading control: Anti-GAPDH.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of SMARCA5 in human cell line MOLT-4 using a SMARCA5 Polyclonal Antibody (Product # PA5-52601).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of SMARCA5 in mouse cell line NIH-3T3 and rat cell line NBT-II using a SMARCA5 Polyclonal Antibody (Product # PA5-52601).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of SMARCA5 was achieved by transfecting HeLa with SMARCA5 specific siRNAs (Silencer® select Product # S16082, S16081). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1%SDS) from the SMARCA5 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SMARCA5 Polyclonal Antibody (Product # PA5-52601, 0.2 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to SMARCA5.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-SMARCA5 Polyclonal Antibody (Product # PA5-52601) and a 130 kDa band corresponding to SMARCA5 was observed across the cell lines tested. Modified whole cell extracts (1%SDS) (30 µg lysate) of HeLa (Lane 1), Jurkat (Lane 2), K-562 (Lane 3), Raji (Lane 4), U-2 OS (Lane 5) and MOLT-4 (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 (0.2 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent staining of SMARCA5 in human cell line A-431 shows positivity in nucleus & nucleoli. Samples were probed using a SMARCA5 Polyclonal Antibody (Product # PA5-52601).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of SMARCA5 was performed using 70% confluent log phase HeLa 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 SMARCA5 Polyclonal Antibody (Product # PA5-52601) at 1 µg/mL 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 dilution), 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 Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of SMARCA5 in human testis using a SMARCA5 Polyclonal Antibody (Product # PA5-52601) shows strong nuclear positivity in cells in seminiferous ducts.

PA5-52601