MA5-17133

antibody from Invitrogen Antibodies

Targeting: NEDD8 Nedd-8

ELISA

Antibody data

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Validation data

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 indirect ELISA, FACS, ICC, IHC, IF and WB applications and shows reactivity with Human samples. The MA5-17133 immunogen is purified recombinant fragment of human NEDD8 expressed in E. Coli. MA5-17133 detects NEDD8 which has a predicted molecular weight of approximately 9kDa.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 1A7
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

NEDD8 protein structure - MA5-17133 shown in red.

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.

Supportive validation

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.

Supportive validation

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).

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.