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Complement Factor Bb 抗体

This anti-Complement Factor Bb antibody is a 小鼠 单克隆 antibody detecting Complement Factor Bb in FACS, ELISA 和 WB. Suitable for 人. Independently validated for use in Western Blotting. This Primary Antibody has been cited in 1 publication.
产品编号 ABIN2473066
发货至: 中国

Quick Overview for Complement Factor Bb 抗体 (ABIN2473066)

抗原

Complement Factor Bb

适用

宿主

  • 6
  • 1
小鼠

克隆类型

  • 6
  • 1
单克隆

标记

  • 2
  • 1
  • 1
  • 1
  • 1
  • 1
This Complement Factor Bb antibody is un-conjugated

应用范围

  • 5
  • 3
  • 3
  • 3
  • 2
  • 2
  • 1
  • 1
Flow Cytometry (FACS), ELISA, Western Blotting (WB)

克隆位点

032B-22-1-X
  • 特异性

    Specificity property: neoantigen
    The monoclonal antibody, clone 032B-22.1.X, binds to a neoantigen expressed on the Bb fragment of human Factor B. This antigen is not expressed on native Factor B and, accordingly, the antibody does not bind to native Factor B found in human plasma.

    产品特性

    Purified IgG

    纯化方法

    Purified

    亚型

    IgG2a
  • 应用备注

    Optimal working dilution should be determined by the investigator.

    说明

    If the Factor B is denatured in any way, the protein can express neoantigen(s) that are not present in functionally active, native Factor B. For example, if the Factor B is bound to a well in a microtiter plate, a portion of it will be conformationally distorted and express the Bb neoantigen. Purification of Factor B can also result in denaturation of a portion of the Factor B and this denatured protein can also express the Bb-specific neoantigen. Finally, lack of careful handling of serum and plasma can result in complement activation and generation of small amounts of Bb and Ba fragments; in this case low levels of Bb will be detectable with this antibody and with the MicroVue assay for Bb.

    限制

    仅限研究用
  • 生效 #103567 (Western Blotting)
    '独立验证'标志
    by
    Complement Physiopathology, Departamento de Microbiología I, Universidad Complutense de Madrid
    No.
    #103567
    日期
    2019.02.22
    抗原
    Bb
    Lot Number
    146096
    Method validated
    Western Blotting
    Positive Control

    Purified Bb fragment and zymosan activated serum

    Negative Control

    Non-activated sera

    Notes

    ABIN2473066 recognizes both Factor B and Bb fragment in plasma samples. It does not specifically recognize a neo-epitope in Bb fragment.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2473066
    Secondary Antibody
    goat anti-mouse Immunoglobulins/HRP (Dako, P0447, lot 20058518)
    Full Protocol
    • Purify Bb fragment from plasma samples.
    • Separate 10ng purified Bb fragment and 0.5µl of zymosan activated serum and 0,5 µl of control serums on a 10% SDS-PAGE electrophoresis gel for 1h 120 V.
    • Activate PVDF membrane with methanol for 15sec and rinse with transfer buffer for 5min. Transfer proteins from the gel to the PVDF membrane using a Mini Trans-Blot cell Tetra Blotting Module (Bio-Rad, 1660827EDU) for 1h at 100V.
    • Stain membrane using Ponceau S to verify protein transfer.
    • Block the membrane with blocking buffer (1x PBS, 3% BSA, 0.1% Tween) for 30min at RT.
    • Incubate membrane with primary mouse anti-complement factor Bb antibody (antibodies-online, ABIN5692886, lot 146096) diluted 1:1000 in working buffer (1x PBS 1% BSA, 0.1% Tween) ON at 4°C.
    • Wash membrane 3x for 5min with PBST (1x PBS, 0.1% Tween).
    • Incubate membrane with secondary goat anti-mouse Immunoglobulins/HRP (Dako, P0447, lot 20058518) diluted 1:1000 in working buffer for 30min at RT.
    • Wash membrane 3x for 5min with PBST.
    • Reveal protein bands using ClarityTM Western ECL substrate (Bio-Rad, 170-5061, lot 102031078) on a ChemiDoc MP Imaging System (Bio-Rad, 17001402).
    Experimental Notes
    • In dentatured samples the complement Factor Bb antibody directed against a neo-epitope in Bb fragment ABIN2473066 reveals two proteins of the expected molecular weight of Factor B (90kDa) and Bb fragment (60kDa). The antibody does not specifically recognize a neo-epitope in Bb fragment and is thus not suitable for selectively detecting the Bb fragment subsequently to denaturation.

    • As an additional control we performed a Western Blot using in-house antibodies which detect Factor B and fragments Bb and Ba (not shown).

  • 状态

    Liquid

    浓度

    1.0 mg/mL
  • Xu, Narayana, Volanakis: "Structural biology of the alternative pathway convertase." in: Immunological reviews, Vol. 180, pp. 123-35, (2001) (PubMed).

  • 抗原

    Complement Factor Bb

    基因ID

    629

    UniProt

    P00751
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