Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Flow cytometry [1]
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Validation data
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- Product number
- PA5-35366 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- EWSR1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with mouse based on sequence homology.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 400 µL
- Concentration
- 0.3 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of EWSR1 in A2058 cell lysate (35 µg/lane) using an EWSR1 polyclonal antibody (Product # PA5-35366).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of EWSR1 was achieved by transfecting A-673 with EWSR1 specific siRNAs (Silencer® select Product # S4887, S4886). Western blot analysis (Fig. a) was performed using whole cell extracts from the EWSR1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with EWSR1 Polyclonal Antibody (Product # PA5-35366, 1:1000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000). 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 EWSR1. A non specific band (*) was observed ~65 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using EWSR1 Polyclonal Antibody (Product # PA5-35366) and a ~80 kDa band corresponding EWSR1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A-673 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), MCF7 (Lane 4), A549 (Lane 5), A-431 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. 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) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A non specific band (*) was observed at ~65 kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of EWSR1 was performed using 70% confluent log phase T-47D 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 EWSR1 Polyclonal Antibody (Product # PA5-35366) at 1:50 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 nuclear and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of EWSR1 showing staining in the nucleus of U-251 cells using an EWSR1 polyclonal antibody (Product # PA5-35366) followed by detection using a fluorescent conjugated secondary antibody (green). Cytoplasmic actin was stained with a fluorescent red phalloidin (7units/mL, 1 h at 37°C). Nuclei were stained with DAPI (blue) (10 µg/mL, 10 min).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of EWSR1 in Hela cells (right) compared to a negative control (left) using an EWSR1 polyclonal antibody (Product # PA5-35366) followed by detection using a FITC-conjugated goat-anti-rabbit secondary antibody.