Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunohistochemistry [2]
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Validation data
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- Product number
- MA5-27439 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- N-WASP Monoclonal Antibody (OTI6A4)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI6A4
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of WASL was achieved by transfecting MCF7 with WASL specific siRNAs (Silencer® select Product # s17133, s17132). Western blot analysis (Fig. a) was performed using whole cell extracts from the WASL knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with N-WASP Monoclonal Antibody (OTI6A4) (Product # MA5-27439, 1:2000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 0.25 µg/mL, 1:4000 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 WASL. An additional band (*) was observed at ~55 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using N-WASP Monoclonal Antibody (OTI6A4) (Product # MA5-27439) and a 60 kDa band corresponding to WASL was observed in SkBr3, SHSY5Y, MCF7, Mouse Lung and Mouse Cerebellum along with an uncharacterized band (*) at ~55 kDa; also, the target protein was absent in U937, HEL92.1.7, HL60, Mouse Kidney and Mouse Skeletal Muscle. Whole cell extracts (30 µg lysate) of SkBr3 (Lane 1), SHSY5Y (Lane 2), MCF7 (Lane 3), U937 (Lane 4), HEL92.1.7 (Lane 5), HL60 (Lane 6), tissue extracts (30 µg lysate) of Mouse Lung (Lane 7), Mouse Cerebellum (Lane 8), Mouse Kidney (Lane 9) and Mouse Skeletal Muscle (Lane 10) were electrophoresed using 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 (1:2000 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
- Immunohistochemistry was performed on paraffin-embedded human kidney tissue. To expose target proteins, heat-induced epitope retrieval by 10mM citric buffer, pH6.0, 100°C for 10min. Following antigen retrieval, tissues were probed with a WASL monoclonal antibody (Product # MA5-27439) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded human spleen tissue. To expose target proteins, heat-induced epitope retrieval by 10mM citric buffer, pH6.0, 100°C for 10min. Following antigen retrieval, tissues were probed with a WASL monoclonal antibody (Product # MA5-27439) at a dilution of 1:500.