PA5-21296

antibody from Invitrogen Antibodies

Targeting: FLOT2 ECS-1, ECS1, ESA, ESA1, M17S1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-21296

Provider

Invitrogen Antibodies

Product name

Flotillin 2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: HeLa, mouse brain, rat brain. Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Flotillin 2 Polyclonal Antibody (Product # PA5-21296). 293T whole cell extracts and 293T exosome extract (2.6 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with Flotillin 2 Polyclonal Antibody (Product # PA5-21296) diluted at 1:250. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flotillin 2 antibody detects Flotillin 2 protein by western blot analysis. Whole cell extracts (30 µg) was separated by 10% SDS-PAGE, and blotted with Flotillin 2 antibody Flotillin 2 Polyclonal Antibody (Product # PA5-21296) diluted by 1:1,000. 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

Flotillin 2 Polyclonal Antibody detects FLOT2 protein by western blot analysis. A. 50 µg mouse brain lysate/extract.10% SDS-PAGE. Flotillin 2 Polyclonal Antibody (Product # PA5-21296) dilution: 1:1,000. 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

Flotillin 2 Polyclonal Antibody detects FLOT2 protein by western blot analysis. A. 50 µg rat brain lysate/extract.10% SDS-PAGE. Flotillin 2 Polyclonal Antibody (Product # PA5-21296) dilution: 1:1,000. 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

Knockdown of FLOT2 achieved by transfecting HeLa with FLOT2 specific siRNAs (Silencer® select Product # s5284, s5285). Western blot analysis (Fig. a) was performed using whole cell extracts from the FLOT2 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Flotillin 2 Polyclonal Antibody (Product # PA5-21296, 1:1000 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 FLOT2.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using anti-Flotillin 2 Polyclonal Antibody (Product # PA5-21296) and a 53 kDa band corresponding to FLOT2 was observed in HeLa and A-431 but not in Molt-4 which is reported to be negative. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2) and Molt-4 (Lane 3) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Flotillin-2 in paraformaldehyde-fixed HeLaS3 cells using a Flotillin-2 polyclonal antibody (Product # PA5-21296) at a 1:100 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of paraformaldehyde-fixed HeLaS3, using Flotillin-2 antibody (Product # PA5-21296) at 1:100 dilution.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human lung cancer, using Flotillin-2 (Product # PA5-21296) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of Flotillin 2 was performed in HeLa whole cell extracts using 5 µg of Flotillin 2 Polyclonal Antibody (Product # PA5-21296). Samples were transferred to a membrane and probed with Flotillin 2 Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.