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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Flow cytometry [1]
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- Product number
- MA5-17133 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NEDD8 Monoclonal Antibody (1A7)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-17133 targets NEDD8 in FACS, ICC, IHC, IF and WB applications and shows reactivity with Human samples.
- Antibody clone number
- 1A7
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NEDD8 using a NEDD8 monoclonal antibody (Product # MA5-17133) against a human NEDD8 (AA: 1-81) recombinant protein.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of NEDD8 was achieved by transfecting HeLa with NEDD8 specific siRNAs (Silencer® select Product # s226684, s9424). Western blot analysis (Fig. a) was performed using whole cell extracts from the NEDD8 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with NEDD8 Monoclonal Antibody (1A7) (Product # MA5-17133, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 0.25µg/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 NEDD8.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti- NEDD8 Monoclonal Antibody (1A7) (Product # PA5-17133) and a 9 kDa band corresponding to NEDD8 was observed across all the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HEK-293 (Lane 2), Jurkat (Lane 3), PC-12 (Lane 4), THP-1 (Lane 5), K-562 (Lane 6), HL-60 (Lane 7), Raw 264.7 (Lane 8), tissue extracts of Rat testis (Lane 9), Mouse brain (Lane 10) and Mouse colon (Lane 11) were electrophoresed Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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, 0.25µg/ml, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NEDD8 using a NEDD8 monoclonal antibody (Product # MA5-17133) against a human NEDD8 (AA: 1-81) recombinant protein.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HeLa cells using NEDD8 monoclonal antibody (Product # MA5-17133) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using NEDD8 monoclonal antibody (Product # MA5-17133) followed with DAB staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of HeLa cells using NEDD8 monoclonal antibody (Product # MA5-17133) (green) and negative control (red).
