- PA5-23086 - Invitrogen Antibodies LAMTOR1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of LAMTOR1 in human liver protein. Sample was incubated in LAMTOR1 polyclonal antibody (Product # PA5-23086).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Ragulator complex protein LAMTOR1 was achieved by transfecting MCF7 with Ragulator complex protein LAMTOR1 specific siRNAs (Silencer® select Product # s29964, s29965). Western Blot analysis (Fig. a) was performed using Membrane enriched extracts from the Ragulator complex protein LAMTOR1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with LAMTOR1 Polyclonal Antibody (Product # PA5-23086, 1:1000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 Ragulator complex protein LAMTOR1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot was performed using Anti-LAMTOR1 Polyclonal Antibody (Product # PA5-23086) and a ~22 kDa band corresponding to Ragulator complex protein LAMTOR1 was observed across cell lines and tissues tested . Membrane enriched extracts (50 µg lysate) of HeLa (Lane 1), Jurkat (Lane 2), NIH/3T3 (Lane 3), U-87 MG (Lane 4), PANC-1 (Lane 5), Mouse Brain (Lane 6), Rat Brain (Lane 7), Rat Kidney (Lane 8) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) 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)
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Experimental details: Immunofluorescent analysis of LAMTOR1 using a polyclonal antibody (Product # PA5-23086).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of LAMTOR1 in HepG2 cells. Samples were incubated in LAMTOR1 polyclonal antibody (Product # PA5-23086) using a dilution of 1:50 followed by FITC (green). Nuclei (Blue) were counterstained with DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Ragulator complex protein LAMTOR1 was performed using 70% confluent log phase Hep G2 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 LAMTOR1 Polyclonal Antibody (Product # PA5-23086) at 1:100 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 along with golgi localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

PA5-23086