Flotillin-2 Recombinant Rabbit Monoclonal Antibody [PSH03-41]
Catalog# HA721986
Flotillin-2 Recombinant Rabbit Monoclonal Antibody [PSH03-41]
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WB
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IHC-P
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IF-Cell
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FC
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Human
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Mouse
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Rat
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Green monkey
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unconjugated
Overview
Product Name
Flotillin-2 Recombinant Rabbit Monoclonal Antibody [PSH03-41]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human Flotillin-2 aa 1-428 / 428.
Species Reactivity
Human, Mouse, Rat, Green monkey
Validated Applications
WB, IHC-P, IF-Cell, FC
Target Molecular Weight
Predicted band size: 47 kDa
Positive Control
HeLa cell lysate, A431 cell lysate, COS-1 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse lung tissue lysate, rat lung tissue lysate, HeLa, 293T cell lysate, A549 cell lysate, MCF7 cell lysate, human lung tissue lysate, mouse skin tissue lysate, rat skin tissue lysate, NIH/3T3, PC-12, human breast cancer tissue, human liver cancer tissue, mouse lung tissue, rat lung tissue.
Conjugation
unconjugated
Clone Number
PSH03-41
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000-1:10,000
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IHC-P
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1:500-1:2,000
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IF-Cell
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1:100
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FC
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1:1,000
Target
Function
Flotillin-2 is a protein that in humans is encoded by the FLOT2 gene. Flotillin 2 (flot-2) is a highly conserved protein isolated from caveolae/lipid raft domains that tether growth factor receptors linked to signal transduction pathways. Flot-2 binds to PAR-1, a known upstream mediator of major signal transduction pathways implicated in cell growth and metastasis, and may influence tumour progression. 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.
Background References
1. Wang W et al. Flotillin 2 Facilitates the Infection of a Plant Virus in the Gut of Insect Vector. J Virol. 2022 Apr
2. Xu Z et al. Flotillin-2 predicts poor prognosis and promotes tumor invasion in intrahepatic cholangiocarcinoma. Oncol Lett. 2020 Mar
Subcellular Location
Cell membrane, Membrane, caveola, Endosome, Membrane.
Synonyms
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
ExpandImages
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Western blot analysis of Flotillin-2 on different lysates with Rabbit anti-Flotillin-2 antibody (HA721986) at 1/5,000 dilution and competitor's antibody at 1/2,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: A431 cell lysate (15 µg/Lane)
Lane 3: COS-1 cell lysate (15 µg/Lane)
Lane 4: NIH/3T3 cell lysate (15 µg/Lane)
Lane 5: PC-12 cell lysate (15 µg/Lane)
Lane 6: Mouse lung tissue lysate (30 µg/Lane)
Lane 7: Rat lung tissue lysate (30 µg/Lane)
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: Lane 1-7 (left): 6 seconds; Lane 1-7 (right): 50 seconds; ECL: K1801;
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 (HA721986) at 1/5,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of Flotillin-2 on different lysates with Rabbit anti-Flotillin-2 antibody (HA721986) at 1/10,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Flotillin-2 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: 10 seconds; ECL: K1801;
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 (HA721986) at 1/10,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling Flotillin-2 with Rabbit anti-Flotillin-2 antibody (HA721986) 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 (HA721986) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Western blot analysis of Flotillin-2 on different lysates with Rabbit anti-Flotillin-2 antibody (HA721986) 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)
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: 20 seconds; ECL: K1801;
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 (HA721986) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of NIH/3T3 cells labeling Flotillin-2 with Rabbit anti-Flotillin-2 antibody (HA721986) 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 (HA721986) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of PC-12 cells labeling Flotillin-2 with Rabbit anti-Flotillin-2 antibody (HA721986) at 1/100 dilution.
Cells were fixed in 80% 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 (HA721986) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Flotillin-2 antibody (HA721986) at 1/2,000 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 (HA721986) at 1/2,000 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. -
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-Flotillin-2 antibody (HA721986) 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 (HA721986) 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. -
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Flotillin-2 antibody (HA721986) 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 (HA721986) 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. -
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Flotillin-2 antibody (HA721986) 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 (HA721986) 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. -
Flow cytometric analysis of HeLa cells labeling Flotillin-2.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721986, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of NIH/3T3 cells labeling Flotillin-2.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721986, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of PC-12 cells labeling Flotillin-2.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721986, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"