Complement Factor Bb anticorps

Détails pour le produit réf. ABIN2473066, Fournisseur: Connectez-vous pour afficher
Clonalité (Clone)
Monoclonal ()
Flow Cytometry (FACS), ELISA, Western Blotting (WB)
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N° du produit (Fournisseur)
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'Independent Validation' signe
Antigène Bb
Numéro du lot 146096
Application validée Western Blotting
Contrôle positif

Purified Bb fragment and zymosan activated serum

Contrôle négative

Non-activated sera


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

Anticorps primaire ABIN2473066
Anticorps secondaire goat anti-mouse Immunoglobulins/HRP (Dako, P0447, lot 20058518)
  • 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).
  • 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).

Validation Images
Western Blotting validation image for anti-Complement Factor Bb antibody (ABIN2473066) Western blot analysis of purified Bb fragment (1), zymosan activated serum (2), and n...
Clone 032B-22-1-X
Isotype IgG2a
Specificité 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.
Attributs du produit Purified IgG
Purification Purified
ID gène 629
UniProt P00751
Indications d'application Optimal working dilution should be determined by the investigator.

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.

Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Produit citée dans: Xu, Narayana, Volanakis: "Structural biology of the alternative pathway convertase." dans: Immunological reviews, Vol. 180, pp. 123-35, 2001 (PubMed).

Background publications Sulser: "[The rhabdomyosarcoma with regard to age, sex, localization, pathological anatomy, and prognosis (author's transl)]." dans: Virchows Archiv. A, Pathological anatomy and histology, Vol. 379, Issue 1, pp. 35-71, 1978 (PubMed).

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