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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunocytochemistry [1]
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- Product number
- MA5-23512 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SMARCA5 Monoclonal Antibody
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Vial size
- 50 µg
- Concentration
- 2 mg/mL
- Storage
- -20°C or -80°C if preferred
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed on nuclear extracts from HeLa cells (HeLa NE, 20 µg) using a SNF2H/SMARCA5 monoclonal antibody (Product # MA5-23512) at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (kDa) is shown on the left), the position of the protein of interest is shown on the right.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed on nuclear extracts from HeLa cells (HeLa NE, 20 µg) using a SNF2H/SMARCA5 monoclonal antibody (Product # MA5-23512) at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (kDa) is shown on the left), the position of the protein of interest is shown on the right.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SMARCA5 Monoclonal Antibody (Product # MA5-23512) and a 130 kDa band corresponding to SMARCA5 was observed in all tested cell line lysates. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), SH-SY5Y (Lane 2), HEK-293 (Lane 3), Jurkat (Lane 4), K-562 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:10000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of SMARCA5 was achieved by transfecting SH-SY5Y with SMARCA5 specific siRNAs (Silencer® select Product # S16081, S16082). Western blot analysis (Fig. A) was performed using Nuclear enriched extracts 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 Monoclonal Antibody (Product # MA5-23512, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. B). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to SMARCA5.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SMARCA5 Monoclonal Antibody (Product # MA5-23512) and a 130kDa band corresponding to SMARCA5 was observed across all cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of Jurkat (Lane 1), K-562 (Lane 2), HeLa (Lane 3), Raji (Lane 4), U-2 OS (Lane 5), Hep G2 (Lane 6) and MOLT-4 (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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)
- Main image
- 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 1% BSA for 1 hour at room temperature. The cells were labeled with SMARCA5 Monoclonal Antibody (Product # MA5-23512) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization to nucleus. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
