WB
Western blot analysis of Flotillin-2 on different lysates with Rabbit anti-Flotillin-2 antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (30 µg/Lane), Lane 2: 293T cell lysate (30 µg/Lane), Lane 3: A549 cell lysate (30 µg/Lane), Lane 4: A431 cell lysate (30 µg/Lane), Lane 5: MCF7 cell lysate (30 µg/Lane), Lane 6: COS-1 cell lysate (30 µg/Lane), Lane 7: NIH/3T3 cell lysate (30 µg/Lane), Lane 8: PC-12 cell lysate (30 µg/Lane), Lane 9: Human lung tissue lysate (30 µg/Lane), Lane 10: Mouse lung tissue lysate (30 µg/Lane), Lane 11: Mouse skin tissue lysate (hot lysis) (30 µg/Lane), Lane 12: Rat skin tissue lysate (30 µg/Lane), Lane 13: Rat lung tissue lysate (30 µg/Lane), Exposure time: 20 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-Flotillin-2 antibody at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Flotillin-2 antibody at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Flotillin-2 antibody at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Flotillin-2 with Rabbit anti-Flotillin-2 antibody at 1/100 dilution. Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Flotillin-2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells labeling Flotillin-2. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Flotillin-2 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human Flotillin-2 aa 1-428 / 428. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:2000-1:10000 IHC:1:500-1:2000 ICC/IF:1:100 FC:1:1000 |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | FLOT2 |
|---|---|
| Gene Synonyms | ESA ECS1 ESA1 ECS-1 M17S1 |
| Gene Full Name | flotillin 2 |
| Gene Summary | Caveolae are small domains on the inner cell membrane involved in vesicular trafficking and signal transduction. This gene encodes a caveolae-associated, integral membrane protein, which is thought to function in neuronal signaling. [provided by RefSeq, Jul 2008] |
| Alternative Names | ECS-1 antibody ECS1 antibody Epidermal surface antigen 1 antibody Epidermal surface antigen antibody ESA antibody ESA1 antibody Flot2 antibody FLOT2_HUMAN antibody Flotillin 2 (epidermal surface antigen 1) antibody Flotillin-2 antibody ECS-1 antibody ECS1 antibody Epidermal surface antigen 1 antibody Epidermal surface antigen antibody ESA antibody ESA1 antibody Flot2 antibody FLOT2_HUMAN antibody Flotillin 2 (epidermal surface antigen 1) antibody Flotillin-2 antibody M17S1 antibody Membrane component chromosome 17 surface marker 1 antibody Membrane component chromosome 17 surface marker 1 homolog antibody membrane component chromosome 17 surface marker 1 (35kD protein identified by monoclonal antibody ECS-1) antibody Membrane component chromosome 17 surface marker 1 antibody REG-1 antibody Reggie-1 antibody Reggie-2 antibody |
| Molecular Weight(MW) | 47kDa |
| Cellular Localization | Cell membrane, Membrane, caveola, Endosome, Membrane. |
WB
Western blot analysis of Flotillin-2 on different lysates with Rabbit anti-Flotillin-2 antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (30 µg/Lane), Lane 2: 293T cell lysate (30 µg/Lane), Lane 3: A549 cell lysate (30 µg/Lane), Lane 4: A431 cell lysate (30 µg/Lane), Lane 5: MCF7 cell lysate (30 µg/Lane), Lane 6: COS-1 cell lysate (30 µg/Lane), Lane 7: NIH/3T3 cell lysate (30 µg/Lane), Lane 8: PC-12 cell lysate (30 µg/Lane), Lane 9: Human lung tissue lysate (30 µg/Lane), Lane 10: Mouse lung tissue lysate (30 µg/Lane), Lane 11: Mouse skin tissue lysate (hot lysis) (30 µg/Lane), Lane 12: Rat skin tissue lysate (30 µg/Lane), Lane 13: Rat lung tissue lysate (30 µg/Lane), Exposure time: 20 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-Flotillin-2 antibody at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Flotillin-2 antibody at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Flotillin-2 antibody at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Flotillin-2 with Rabbit anti-Flotillin-2 antibody at 1/100 dilution. Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Flotillin-2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling Flotillin-2. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:2000-1:10000 IHC:1:500-1:2000 ICC/IF:1:100 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
