PA5-28105

antibody from Invitrogen Antibodies

Targeting: TICAM1 MGC35334, PRVTIRB, TICAM-1, TRIF

Antibody data

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Validation data

TICAM1 protein structure - PA5-28105 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using TRIF Polyclonal Antibody (Product # PA5-28105). Sample (30 µg of whole cell lysate). Lane A: Hela . 7.5% SDS PAGE. TRIF Polyclonal Antibody (Product # PA5-28105) diluted at 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of TIR domain-containing adapter molecule 1 was achieved by transfecting Raji with TIR domain-containing adapter molecule 1 specific siRNAs (Silencer® select Product # S531859, S45114). Western blot analysis (Fig. a) was performed using Whole cell extracts from the TIR domain-containing adapter molecule 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TRIF Polyclonal Antibody (Product # PA5-28105, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 TIR domain-containing adapter molecule 1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-TRIF Polyclonal Antibody (Product # PA5-28105) and a 76.4 kDa band corresponding to TIR domain-containing adapter molecule 1 was observed across in Raji and Ramos, but not in HL-60 and HEL 92.1.7. Whole cell extracts (40 µg lysate) of Raji (Lane 1), HL-60 (Lane 2), Ramos (Lane 3), HEL 92.1.7 (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: TRIF Polyclonal Antibody detects TRIF protein at cytosol by immunofluorescent analysis. Sample: A431 cells were fixed in 4% paraformaldehyde for 10 min. Green: TRIF protein stained by TRIF Polyclonal Antibody (Product # PA5-28105) diluted at 1:100. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of TIR domain-containing adapter molecule 1 was performed using 70% confluent log phase Ramos 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 TRIF Polyclonal Antibody (Product # PA5-28105) at 1/200 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 cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of TIR domain-containing adapter molecule 1 was performed using 70% confluent log phase Raji 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 TRIF Polyclonal Antibody (Product # PA5-28105) at 1:200 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 dilution), 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 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents HEL 92.1.7 cells, showing no expression of TICAM1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.