Recombinant Mouse Anti-C. burnetii PI-LPS Antibody (1E4) (CAT#: FAMAB-0041CQ)

• Immunogen
• C. burnetii Nine Mile PI antigen

• Host Species
• Mouse

• Type
• Mouse IgG2a

• Specificity
• C. burnetii phase I lipopolysaccharide (PI-LPS)

• Species Reactivity
• C. burnetii

• Clone
• 1E4

• Applications
• ELISA, WB, Inhib, IF, FuncS

• Related Disease
• Q fever

• Purity
• >95% as determined by analysis by SDS-PAGE

• Storage
• Store at -20°C for long-term storage. Avoid freeze/thaw cycles.

• Application Notes
• Enzyme-linked Immunosorbent Assay was performed as modified from the method described previously and used to screen hybridoma supernatants for their reactivity with PI and PII Ags and isotype-cloned hybridomas. One hundred microliters of inactivated NMI or NMII Ag at 0.5 μg/ml in 0.05 M carbonate/bicarbonate coating buffer (pH 9.6) was added to each well of a 96-well microtiter plate and coated at 4˚C for 24 h. The plates were blocked with 1% BSA in PBST buffer (0.05% Tween 20 in PBS) and then incubated with 100 μl hybridoma supernatants or diluted purified mAb at 37˚C for 2 h. After washing five times with PBST buffer, the plates were incubatd with 100 μl HRP-conjugated goat anti-mouse IgM, IgG, IgG1, IgG2a, IgG2b, or IgG3 (1:1000 dilution) at 37˚C for 2 h. After washing five times with PBSTbuffer, the Sigma Fast O-Phenylenediamine Dihydrochloride Tablet Sets (Sigma-Aldrich) were used as substrates, and the absorbance was measured at 490 nm by the SPECTRA MAX M2 system using SoftMax program (Molecular Devices, Sunnyvale, CA).
  Western blot
  NMI and NMII whole-cell Ags were separated by SDS-PAGE and transferred electrophoretically to nitrocellulose membranes in Tris-glycine
  buffer. The membranes were blocked for 1 h at room temperature in TBS buffer with 0.05% Tween 20 (TBST) and 5% nonfat dry milk, and then
  incubated with diluted mAb or immune serum at 4˚C overnight. After washing five times (5 min each wash) with TBST buffer, the membranes
  were incubated with HRP-conjugated goat anti-mouse IgG (1:10,000 dilution) for 1 h at room temperature. The reactions were detected by using an ECL Western blot detection kit (Amersham Pharmacia, Piscataway, NJ). To determine whether purified 1E4 or immune sera recognize protein or LPS, the reactivity of 1E4 and immune sera to proteinase K-treated and untreated PI and PII Ags was also tested by Western blotting. Digestion of PI and PII whole-cell Ags by proteinase K was performed.

• Alternative Names
• C. burnetii PI-LPS; C. burnetii phase I lipopolysaccharide

• FAMAB-0041CQ-F(E)
• Recombinant Mouse Anti-C. burnetii PI-LPS Antibody Fab Fragment (1E4)

• FAMAB-0041CQ-S(P)
• Recombinant Mouse Anti-C. burnetii PI-LPS Antibody scFv Fragment (1E4)

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