Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [6]
- Flow cytometry [2]
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- Product number
- PA5-79268 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Flotillin 2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 500 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Flotillin 2 in Lane 1: rat liver tissue lysate, Lane 2: rat lung tissue lysate, Lane 3: mouse liver tissue lysate, Lane 4: mouse lung tissue lysate, Lane 5: mouse testis tissue lysate, Lane 6: human placenta tissue lysate, Lane 7: A375 whole cell lysate, Lane 8: HeLa whole cell lysate, Lane 9: 22RV1 whole cell lysate, Lane 10: NIH3T3 whole cell lysate using 40-50 µg per well. Sample was incubated with Flotillin 2 (Product # PA5-79268) at a dilution of 0.5 µg/mL.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of Flotillin 2 in, Lane 1: human 293T whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human A375 whole cell lysates, Lane 4: human A549 whole cell lysates, Lane 5: rat thymus tissue lysates, Lane 6: rat lung tissue lysates, Lane 7: mouse thymus tissue lysates, Lane 8: mouse lung tissue lysates. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. The membrane was blocked with 5% non-fat milk/TBS for 1. 5 hour at RT. The membrane was incubated with Flotillin 2 Polyclonal Antibody (Product # PA5-79268) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for Flotillin 2 at approximately 47-50 kDa. The expected band size for Flotillin 2 is at 47-50 kDa.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Knockdown of FLOT2 was 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 HeLa 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 # PA5-79268, 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.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using anti-Flotillin 2 Polyclonal Antibody (Product # PA5-79268) and a 53 kDa corresponding to FLOT2 was observed in HeLa, A-431, NIH/3T3, L6, Mouse Brain, Rat Brain and Mouse Kidney and a 47 kDa band corresponding to FLOT2 was observed in Mouse Brain. The protein expression was not found in Molt-4 and Mouse skeletal Muscle. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), Molt-4 (Lane 3), NIH/3T3 (Lane 4), L6 (Lane 5), tissue extracts (30 µg lysate) of Mouse Brain (Lane 6), Rat Brain (Lane 7), Mouse Kidney (Lane 8) and Mouse Skeletal Muscle (Lane 9) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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).
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemistry analysis of Flotillin 2 using anti-Flotillin 2 antibody (Product # PA5-79268). Flotillin 2 was detected in a section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-Flotillin 2 antibody (Product # PA5-79268)overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Flotillin 2 on paraffin-embedded rat kidney tissue. Sample was incubated with Flotillin 2 polyclonal antibody (Product# PA5-79268).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Flotillin 2 on paraffin-embedded human mammary cancer tissue. Sample was incubated with Flotillin 2 polyclonal antibody (Product# PA5-79268).
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry analysis of Flotillin 2 on rat small intestine tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with Flotillin 2 polyclonal antibody (Product# PA5-79268) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Flotillin 2 on paraffin-embedded mouse intestine tissue. Sample was incubated with Flotillin 2 polyclonal antibody (Product# PA5-79268).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Flotillin 2 on rat kidney tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with Flotillin 2 polyclonal antibody (Product# PA5-79268) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Flotillin 2 on mouse kidney tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with Flotillin 2 polyclonal antibody (Product# PA5-79268) with a dilution of 1 µg/mL (overnight at 4°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry of Flotillin 2 in RH-35 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with Flotillin 2 Polyclonal Antibody (Product # PA5-79268) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry of Flotillin 2 in U2OS cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with Flotillin 2 Polyclonal Antibody (Product # PA5-79268) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.