Anti-Flotillin 1 抗体 [EPR6041] (ab133497)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6041] to Flotillin 1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Flotillin 1 antibody [EPR6041]
Flotillin 1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR6041] to Flotillin 1 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Flotillin 1 aa 400-500 (C terminal). The exact sequence is proprietary.
Database link: O75955 -
ポジティブ・コントロール
- WB: HEK293T, HAP1, K562, A431, Jurkat, and HeLa cell lysates; Mouse brain and kidney lysates; Rat kidney lysate. IP: K562 cell lysate. IHC-P: Human cervical carcinoma tissue. Flow Cyt (intra): Jurkat cells. ICC/IF: Jurkat cells.
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR6041 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab133497の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
1/50.
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WB | (4) |
1/10000 - 1/50000. Detects a band of approximately 48 kDa (predicted molecular weight: 47 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP |
1/10 - 1/100.
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ICC/IF |
1/500.
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特記事項 |
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Flow Cyt (Intra)
1/50. |
WB
1/10000 - 1/50000. Detects a band of approximately 48 kDa (predicted molecular weight: 47 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
1/10 - 1/100. |
ICC/IF
1/500. |
ターゲット情報
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機能
May act as a scaffolding protein within caveolar membranes, functionally participating in formation of caveolae or caveolae-like vesicles. -
配列類似性
Belongs to the band 7/mec-2 family. Flotillin subfamily. -
細胞内局在
Cell membrane. Membrane > caveola. Melanosome. Endosome. Membrane-associated protein of caveolae. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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参照データベース
- Entrez Gene: 10211 Human
- Entrez Gene: 14251 Mouse
- Entrez Gene: 64665 Rat
- Omim: 606998 Human
- SwissProt: O75955 Human
- SwissProt: O08917 Mouse
- SwissProt: Q9Z1E1 Rat
- Unigene: 179986 Human
see all -
別名
- FLOT 1 antibody
- FLOT1 antibody
- FLOT1_HUMAN antibody
see all
画像
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Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab133497 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunofluorescence staining of Jurkat cells with purified ab133497 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab133497 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : FLOT1 knockout HEK293T cell lysate
Lane 3 : HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab133497 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.
ab133497 Anti-Flotillin 1 antibody [EPR6041] was shown to specifically react with Flotillin 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267276 (knockout cell lysate ab257109) was used. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. ab133497 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ab133497 (purified) at 1/60 immunoprecipitating Flotillin 1 in 10 μg K562 (Lanes 1 and 2, observed at 48 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling Flotillin 1 with purified ab133497 at 1/50 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr®488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
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Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Flotillin 1 using unpurified ab133497 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Flotillin 1 knockout HAP1 whole cell lysate (20 µg)Lanes 1 - 2: Merged signal (red and green). Green - ab133497 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab133497 was shown to specifically react with Flotillin 1 in wild-type HAP1 cells as signal was lost in Flotillin 1 knockout cells. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. Ab133497 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/10000 dilution (purified)
Lane 1 : K562 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 47 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/10000 dilution (purified)
Lane 1 : mouse brain lysate
Lane 2 : mouse kidney lysate
Lane 3 : rat kidney lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L)
Predicted band size: 47 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/10000 dilution
Lane 1 : K562 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 47 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (60)
ab133497 は 60 報の論文で使用されています。
- Byappanahalli AM et al. Mitochondrial DNA and inflammatory proteins are higher in extracellular vesicles from frail individuals. Immun Ageing 20:6 (2023). PubMed: 36710345
- You B et al. IGFBP2 derived from PO-MSCs promote epithelial barrier destruction by activating FAK signaling in nasal polyps. iScience 26:106151 (2023). PubMed: 36866245
- Huang Y et al. Cellular cholesterol loss by DHCR24 knockdown leads to Aβ production by changing APP intracellular localization. J Lipid Res 64:100367 (2023). PubMed: 37011864
- Palamà MEF et al. Xeno-free cultured mesenchymal stromal cells release extracellular vesicles with a "therapeutic" miRNA cargo ameliorating cartilage inflammation in vitro. Theranostics 13:1470-1489 (2023). PubMed: 37056573
- Carvalho-Rosa JD et al. Epileptiform activity influences theta-burst induced LTP in the adult hippocampus: a role for synaptic lipid raft disruption in early metaplasticity? Front Cell Neurosci 17:1117697 (2023). PubMed: 37228704