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Flotillin 1 抗体 (C-Term)

FLOT1 适用: 人, Cow, 犬, Pig WB, IHC (p) 宿主: 山羊 Polyclonal unconjugated
产品编号 ABIN374222
发货至: 中国
  • 抗原 See all Flotillin 1 (FLOT1) 抗体
    Flotillin 1 (FLOT1)
    抗原表位
    • 8
    • 7
    • 6
    • 4
    • 4
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    C-Term
    适用
    • 58
    • 26
    • 25
    • 7
    • 6
    • 6
    • 5
    • 4
    • 4
    • 3
    • 3
    • 1
    • 1
    • 1
    人, Cow, 犬, Pig
    宿主
    • 56
    • 2
    • 1
    山羊
    克隆类型
    • 48
    • 11
    多克隆
    标记
    • 28
    • 7
    • 4
    • 4
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    This Flotillin 1 antibody is un-conjugated
    应用范围
    • 37
    • 23
    • 10
    • 7
    • 4
    • 3
    • 2
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
    特异性
    This antibody detetcs (FLOT1) at C-term.
    交叉反应 (详细)
    Species reactivity (expected):Dog, Pig, Cow.
    Species reactivity (tested):Human.
    纯化方法
    Affinity chromatography
    免疫原
    ptide with sequence C-SISQVNHKPLRTA, from the C Terminus of the protein sequence Genename: FLOT1
    Top Product
    Discover our top product FLOT1 Primary Antibody
  • 应用备注
    Peptide ELISA: 1/32000. Western blot: 0,3 - 1,0 1 μg/mL. Approx 48 kDa band observed in H460 lysates (predictedMW of 48 kDa according to NP_005794).
    Other applications not tested.
    Optimal dilutions are dependent on conditions and should be determined by the user.
    限制
    仅限研究用
  • 生效 #101253 (Western Blotting)
    '独立验证'标志
    by
    Internal Disease Clinic, Faculty of Veterinary Medicine, University of Zagreb
    No.
    #101253
    日期
    2017.05.03
    抗原
    Flotillin-1
    Lot Number
    Method validated
    Western Blotting
    Positive Control
    extracellular vesicles from bovine follicular fluid, verified with CD63 antibody
    Negative Control
    non-enriched fractions
    Notes
    Passed. The flotillin-1 antibody ABIN374222 specifically reveals a band of the expected molecular weight in fractions enriched for exosomes.
    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN374222
    Secondary Antibody
    anti-goat-HRP
    Full Protocol
    • Isolate extracellular vesicles from 0.5ml follicular fluid from cow obtained from abattoir using the qEV size-exclusion chromatography kit (iZON Science, qEVoriginal Size Exclusion Columns, NC0888135):
      • Equilibrate columns with 10ml PBS.
      • Apply the follicular fluid.
      • Retain the first 3ml of the eluate as void volume negative control (V0).
      • Collect the next three 0.5ml fractions (1, 2, 3) which are expected to contain extracellular vesicles.
      • Collect next 0.5ml fraction (4) which is expected to contain proteins but not extracellular vesicles from follicular fluid.
    • Concentrate fractions 25x using Microcon filters (Millipore, Ultracel YM-30, MRCF0R30) with a 25kDa cut-off.
    • Boil all five fractions and 1µl of untreated follicular fluid as positive control in 1x Laemmli SDS sample buffer for 5min.
    • Separate all protein in each fraction, 10µg for fraction 2, the richest in protein on a freshly cast 4% acrylamide stacking gel and 10% acrylamide separation SDS-PAGE gel.
    • Transfer proteins onto a nitrocellulose membrane (GE Healthcare,Amersham Protran 0.45µm, 10600002) in an electroblotter tank (Biostep, electroblotting module GV100- EBGRM) Western blotting system using 20% methanol-containing transfer buffer for 2h at 150mA at RT.
    • Block the membrane with Odyssey blocking buffer (PBS) (Odyssey, 927-400000) for 1h at RT.
    • Incubation with primary goat anti-flotillin 1 antibody (antibodies-online, ABIN374222) diluted 1:500 in blocking buffer ON at 4°C.
    • Wash membrane 3x 5min with TBST (TBS, 0.1% Tween-20).
    • Incubate with secondary anti-goat-HRP antibody diluted 1:5000 in blocking buffer for 1h at RT.
    • Wash membrane 3x 5min with TBST.
    • Incubate membrane in Luminol for (Santa Cruz Biotechnology Inc., ImmunoCruz western blotting luminol reagent, SC-2048) 5min at RT.
    • Reveal chimiuminescent bands using a chemiluminescence detector (LI-COR, Odyssey Fc, OFC-0966) imaging system.
    • Strip membranes usingmild stripping protocol:
      • Incubate membrane with stripping buffer (1.5% (w/v) glycine, 0.0% (w/v) SDS, 1% (v/v) tween 20, pH2.2) shaking for 2x 10min at RT.
      • Wash membrane 2x 10min with PBS.
      • Wash membrane2x 5min in TBS.
      • Block the membrane with blocking buffer.
    • Incubate with primary goat anti-CD63 antibody (antibodies-online, ABIN1440014, lot 0047080912) diluted 1:200 in blocking buffer ON at 4°C.
    • Repeat the same steps as for the goat anti-flotillin 1 antibody.
    Experimental Notes
    We observed an albumin band in western blot of some fractions when we used a different secondary (HRP conjugated mouse anti-CD9 antibody) (not shown). Albumin was not observed with ABIN374222.
  • 浓度
    0.5 mg/mL
    缓冲液
    Tris saline, pH ~7.3, 0.02 % Sodium Azide, 0.5 % BSA
    储存液
    Sodium azide
    注意事项
    This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    注意事项
    Avoid repeated freezing and thawing.
    储存条件
    4 °C/-20 °C
    储存方法
    Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
  • 抗原
    Flotillin 1 (FLOT1)
    别名
    Flotillin-1 / FLOT1 (FLOT1 产品)
    别名
    reggie-2 antibody, flot1 antibody, Rareg antibody, flot1c antibody, flotillin-1 antibody, flotillin1 antibody, flot1a antibody, reggie2 antibody, si:dz44o19.1 antibody, wu:fj45g11 antibody, flotillin 1 antibody, flotillin-1 antibody, putative flotillin-1 antibody, flotillin 1 S homeolog antibody, flotillin 1 L homeolog antibody, flotillin 1a antibody, FLOT1 antibody, Flot1 antibody, flot1 antibody, LOC5566722 antibody, CpipJ_CPIJ008623 antibody, Smp_016200.1 antibody, flot1.S antibody, flot1.L antibody, flot1a antibody
    背景
    Flotillin-1 is a lipid raft-associated protein that has been implicated in various cellular processes: it has been reported to function as a molecular link between lipid rafts of the plasma membrane and a multimeric signaling complex at the actin cytoskeleton, to associate with caveolae , regulating vesicular trafficking and signal transduction, and to have mitogenic properties. Its subcellular localisation seems to be cell-type-specific. It has been found associated with the cell membranes, nucleus, cytoplasm and different cytoplasmic organelles.Synonyms: REG-2, Reg2, Reggie-2
    基因ID
    10211
    NCBI登录号
    NP_005794
    UniProt
    O75955
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