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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
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- Product number
- MA1-20168 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- FADD Monoclonal Antibody (FD19)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- Recommended positive controls: A431.
- Antibody clone number
- FD19
- Concentration
- 2.1 mg/mL
Submitted references Induction of interferon-β and interferon signaling by TRAIL and Smac mimetics via caspase-8 in breast cancer cells.
Granqvist V, Holmgren C, Larsson C
PloS one 2021;16(3):e0248175
PloS one 2021;16(3):e0248175
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of FADD was achieved by transfecting MDA-MB-231 with FADD specific siRNAs (Silencer® select Product # S16705, S16706). Western blot analysis (Fig. a) was performed using Whole cell extracts from the FADD knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with FADD Monoclonal Antibody (FD19) (Product # MA1-20168, 1ug/mL ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:2000). 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 FADD.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-FADD Monoclonal Antibody (FD19)(Product # MA1-20168) and a 23kDa band corresponding to FADD was observed across tested Whole cell extracts (30 µg lysate) of HT-29(Lane 1), HT-29(d-actinomycin treated) (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000) 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
- Fig 6 Involvement of FADD in TRAIL-mediated IFN signaling. A-D) MCF-7 cells were stimulated with 100 ng/ml TRAIL for 2 h in the presence of 20 muM zVAD-FMK and lysates were immunoprecipitated with antibodies towards FADD (A, B) or caspase-8 (C, D). Lysates and immunoprecipitates were subjected to Western blot for FADD and caspase-8 (A and C). Intensities in bands for the immunoprecipitates were quantified (B and D). E-G) MCF-7 cells were transfected for 72 h with a combination of two siRNAs targeting FADD (E) followed by treatment with TRAIL (100 ng/ml) for 48h. Lysates were subjected to Western blot with indicated antibodies (F) and intensities in bands were quantified (G). Data (B,D,G) are mean +- SEM with individual data points indicated and experiments were performed three (B) or four (D,G) times. * denotes p
