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Anti-Flotillin 1 FLOT1 Rabbit Monoclonal Antibody

Figure 1. Western blot analysis of Flotillin 1 using anti-Flotillin 1 antibody (M05980).
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 ug of sample under reducing conditions.
Lane 1: human U2OS whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: human PC-3 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat lung tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse lung tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Flotillin 1 antigen affinity purified monoclonal antibody (Catalog # M05980) at 1:500 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 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Flotillin 1 at approximately 47 kDa. The expected band size for Flotillin 1 is at 47 kDa.
Figure 2. IHC analysis of Flotillin 1 using anti-Flotillin 1 antibody (M05980).
Flotillin 1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-Flotillin 1 Antibody (M05980) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 3. IHC analysis of Flotillin 1 using anti-Flotillin 1 antibody (M05980).
Flotillin 1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-Flotillin 1 Antibody (M05980) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 4. IHC analysis of Flotillin 1 using anti-Flotillin 1 antibody (M05980).
Flotillin 1 was detected in a paraffin-embedded section of human lung squamous cell carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-Flotillin 1 Antibody (M05980) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IP
Primary Accession	O75955
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-Flotillin 1 FLOT1 Rabbit Monoclonal Antibody . Tested in WB, IHC, IP applications. This antibody reacts with Human, Mouse, Rat.

Additional Information
Gene ID	10211
Other Names	Flotillin-1, FLOT1
Calculated MW	47355 Da
Application Details	WB 1:500-1:2000 IHC 1:50-1:200 IP 1:50
Subcellular Localization	Cell membrane; Peripheral membrane protein. Membrane, caveola; Peripheral membrane protein. Melanosome. Endosome. Membrane-associated protein of caveolae. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Source	Eukaryota
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: BHB-8
Immunogen	A synthesized peptide derived from human Flotillin 1
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	FLOT1
Function	May act as a scaffolding protein within caveolar membranes, functionally participating in formation of caveolae or caveolae-like vesicles.
Cellular Location	Cell membrane; Peripheral membrane protein. Endosome Membrane, caveola {ECO:0000250\|UniProtKB:O08917}; Peripheral membrane protein {ECO:0000250\|UniProtKB:O08917}. Melanosome. Membrane raft. Note=Identified by mass spectrometry in melanosome fractions from stage I to stage IV (PubMed:17081065) Membrane-associated protein of caveola (By similarity) {ECO:0000250\|UniProtKB:O08917, ECO:0000269\|PubMed:17081065}

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol