- PA5-78238 - Invitrogen Antibodies RAB7A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of RAB7A in whole cell lysate using 30 µg of protein. Samples were separated with 12% SDS-PAGE and incubated with RAB7A polyclonal antibody (Product # PA5-78238) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of RAB7A in whole cell lysate using 30 µg of protein. Samples were separated with 12% SDS-PAGE and incubated with RAB7A polyclonal antibody (Product # PA5-78238) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using RAB7A Polyclonal Antibody (Product # PA5-78238). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with Rab7 antibody RAB7A Polyclonal Antibody (Product # PA5-78238) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using RAB7A Polyclonal Antibody (Product # PA5-78238). Non-transfected (–) and transfected (+) 293T whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with RAB7A Polyclonal Antibody (Product # PA5-78238) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Ras-related protein Rab-7a was achieved by transfecting A549 with Ras-related protein Rab-7a specific siRNAs (Silencer® select Product # S15443, S15444 ). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Ras-related protein Rab-7a knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RAB7A Polyclonal Antibody (Product # PA5-78238, 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 Ras-related protein Rab-7A.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-RAB7A Polyclonal Antibody (Product # PA5-78238) and a 23kDa band corresponding to Ras-related protein Rab-7a was observed across cell lines. Whole cell extracts (30 µg lysate) of A549 (Lane 1), MCF7 (Lane 2), NIH/3T3 (Lane 3), HeLa (Lane 4), Mouse Heart (Lane 5), Rat Heart (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (1:1000) 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: RAB7A Polyclonal Antibody detects Rab7A protein at cytoplasm by immunofluorescent analysis. Sample: A431 cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Rab7A protein stained by RAB7A Polyclonal Antibody (Product # PA5-78238) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of RAB7A was performed in DIV9 rat E18 primary cortical neuron cells fixed in 4% paraformaldehyde at RT for 15 min. Green: RAB7A Polyclonal Antibody (Product # PA5-78238) diluted at 1:500. Red: beta Tubulin 3/ Tuj1, stained by beta Tubulin 3/ Tuj1 antibody. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of Ras-related protein Rab-7a was performed using 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with RAB7A Polyclonal Antibody (Product # PA5-78238) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (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)
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Experimental details: Rab7 antibody detects Rab7 protein at cytoplasm in mouse liver by immunohistochemical analysis. Sample: Paraffin-embedded mouse liver. Rab7 antibody (Product # PA5-78238) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Rab7 antibody detects Rab7 protein at cytoplasm in rat intestine by immunohistochemical analysis. Sample: Paraffin-embedded rat intestine. Rab7 antibody (Product # PA5-78238) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

PA5-78238