- PA5-31674 - Invitrogen Antibodies RAB35 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of RAB35 using 30 µg of U87-MG lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a RAB35 polyclonal antibody (Product # PA5-31674) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of RAB35 was performed by separating 30 µg of various whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a RAB35 Polyclonal Antibody (Product # PA5-31674) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of RAB35 was performed by separating 50 µg of mouse tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a RAB35 Polyclonal Antibody (Product # PA5-31674) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using RAB35 Polyclonal Antibody (Product # PA5-31674). Mouse tissue extract (50 µg) was separated by 12% SDS-PAGE, and the membrane was blotted with RAB35 Polyclonal Antibody (Product # PA5-31674) diluted at 1:1,000. 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 RAB35 was achieved by transfecting Caki-1 with RAB35 specific siRNAs (Silencer® select Product # SS21707, SS21708). Western blot analysis (Fig. a) was performed using whole cell extracts from the RAB35 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RAB35 Polyclonal Antibody (Product # PA5-31674, 1:2000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) and detected by chemiluminescence using the iBright™ FL1500 Imaging System (Product # A44115). 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 RAB35.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of RAB35 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a RAB35 Polyclonal Antibody (Product # PA5-31674) at a dilution of 1:2000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using RAB35 Polyclonal Antibody (Product # PA5-31674) and a 23 kDa band corresponding to RAB35 was observed across cell lines and tissue tested. Whole cell extracts (30 µg lysate) of Raji (Lane 1), Jurkat (Lane 2), Caki-1 (Lane 3), 769-P (Lane 4), Mouse Cerebellum (Lane 5) and Mouse Lung (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. 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) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of RAB35 was performed in U87-MG cells fixed in 4% paraformaldehyde at RT for 15 min. Green: RAB35 Polyclonal Antibody (Product # PA5-31674) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded human breast cancer, using RAB35 (Product # PA5-31674) antibody at 1:250 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

PA5-31674