Borrelia Burgdorferi 抗体

ABIN236885 产品详细信息, 供应商: Log in to see
抗原
适用
Borrelia burgdorferi
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宿主

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克隆类型
多克隆
标记
非结合性
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应用范围
Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
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选项
Supplier
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Supplier Product No.
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'独立验证'标志
抗原 Borrelia Burgdorferi
Lot Number 10L35615
Method validated Immunofluorescence
Positive Control Macrophages expressing a protein X locating at plasma and endomembranes, infected with Borrelia burgdorferi B31.
Negative Control Macrophages expressing a protein X locating at plasma and endomembranes.
Notes Passed. ABIN236885 has been shown to specifically label Borrelia burgdorferi B31 inside human macrophages and extracellularly.
Primary Antibody ABIN236885
Secondary Antibody anti-rabbit-AF488 (Jackson ImmunoResearch Inc, 111-545-003), anti-rabbit-Alexa568 (Thermo Fisher Scientific, A11011)
实验流程
  • Transfection and infection:
    • Isolate human peripheral blood monocytes from buffy coats and differentiate them into macrophages in RPMI growth media with 20% human serum and 5% L-Glutamin-Pen/Strep (Sigma-Aldrich, G6784) at 37°C, 5% CO2.
    • Transfect primary human macrophages with an expressing plasmid for a protein X that binds to plasma membranes and endomembranes using Neon Transfection System (Thermo Fisher Scientific, MPK5000) for 3h at 37°C and seeded on 12mm glass coverslips (see http://www.ncbi.nlm.nih.gov/pubmed/24696301).
    • Infect macrophages expressing protein X with Borrelia burgdorferi B31 for 1h at 37°C and MOI 100.
    • Fix cells 10 min with 4% methanol-free formaldehyde.
  • Extracellular immunofluorescence staining.
    • Block with 40µl of 2% BSA/PBS containing 5% normal human serum (Jackson ImmunoResearch Inc, 009-000-001) and 5% normal goat serum (Jackson ImmunoResearch Inc, 005-000-001) for 30min at RT.
    • Wash 2x with PBS.
    • Incubation with 40µl primary rabbit-anti-Borrelia burgdorferi antibody (antibodies-online, ABIN236885, lot 10L35615) diluted 1:1000 in PBS for 30min at RT.
    • Wash 2x with PBS.
    • Incubation with 40µl secondary anti-rabbit-AF488 antibody (Jackson ImmunoResearch Inc, 111-545-003) diluted 1:200 in PBS for 30min at RT.
    • Wash 2x with PBS.
  • Intracellular immunofluorescence staining:
    • Permeabilize cells with 0.05% Triton X-100/PBS for 10 min at RT.
    • Wash 2x with PBS.
    • Incubation with 40µl primary rabbit-anti-Borrelia burgdorferi antibody (antibodies-online, ABIN236885, lot 10L35615) diluted 1:1000 in PBS for 30min at RT.
    • Wash 2x with PBS.
    • Incubation with 40µl of anti-rabbit-AF568 antibody (Thermo Fisher Scientific, A11011) diluted 1:200 for 30 min at RT.
    • Wash 2x with PBS.
    • Mount coverslips with 6µl prewarmed Mowiol 4-88 mounting medium (Sigma-Aldrich, 81381) on object slides for at least 1h.
  • Image acquisition:
    • Image acquisition was performed with a Leica TCS SP5 (with a Leica TCS SP5 AOBS confocal point scanner), equipped with an oil-immersion HCX PL APO 63x NA 1.4 λblue objective.
Validation Images
Immunofluorescence validation image for anti-Borrelia Burgdorferi antibody (ABIN236885) Staining of a human macrophage after infection with Borrelia burgdorferi B31. ...
免疫原 Whole Cell preparation from B. burgdorferi Strain: B31 ATCC 35210
特异性 Reactive Western blot bands at 83kD, 41kD, 34kD and 31kD and additional low MW bands. Cross-reacts with Treponema pallidum, B. hermsii and B. parkerii.
产品特性 Rabbit anti B. burgdorferi, Rabbit antibody to Borrelia burgdorferi
纯化方法 Protein A Chromatography. Ascites
纯度 > 90 % pure
研究领域 Bacteria
应用备注 Suitable for use in IFA, Western blot and immunohistochemistry (formalin fixed/paraffin). IFA specimens should be acetone-fixed for best results. Also suitable for conjugation purposes. Each laboratory should determine an optimum working titer for use in its particular application. Other applications have not been tested but use in such assays should not necessarily be excluded.
限制 仅限研究用
状态 Liquid
浓度 4-5 mg/mL
缓冲液 0.01 M PBS, pH 7.2 Product contains no stabilizing proteins, 0.1 % Sodium azide
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 multiple freeze/thaw cycles.
Centrifuge product if not completely clear after standing at room temperature.
Prepare working dilution only prior to immediate use.
储存条件 4 °C/-20 °C
储存方法 Short-term (up to 6 months) store at 2-8 °C. Long term, aliquot and store at -28 °C.
有效期 6 months
有引用在: Naj, Linder: "ER-Coordinated Activities of Rab22a and Rab5a Drive Phagosomal Compaction and Intracellular Processing of Borrelia burgdorferi by Macrophages." in: Cell reports, Vol. 12, Issue 11, pp. 1816-30, 2015 (PubMed).

Hoffmann, Naj, Linder: "Daam1 is a regulator of filopodia formation and phagocytic uptake of Borrelia burgdorferi by primary human macrophages." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 28, Issue 7, pp. 3075-89, 2014 (PubMed).

背景 Pereira, Marques: "[Analysing the content of the section 'Students' page' of the Annaes de Enfermagem]" in: Revista brasileira de enfermagem, Vol. 59 Spec No, pp. 428-31, 2006 (PubMed).

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