To assess IgM deposition, 1 × 107R. parkeri or R. australis bacilli were fixed in 4% PFA solution. After washing with PBS-glycine, bacteria were incubated with 30% of NHS, IgM-dplHS, or PBS for 1 hr at RT or overnight at 4°C. To assess both human IgM and C1q deposition on Rickettsia bacilli, all samples were treated with rabbit anti-human C1q (Thermofisher), Alexa Fluor-546-conjugated donkey anti-rabbit-IgG (Invitrogen) and AlexaFluor-488-conjugated goat IgG anti-human IgM (Invitrogen). Mouse IgM was measured as above but with C57BL6/J serum and AlexaFluor-488-conjugated anti-mouse IgM (clone AF6–78, Invitrogen). Fluorescence was assessed on a FACSCelesta Cell Analyzer (BD Biosciences) using fluorescein isothiocyanate (FITC) and phycoerythrin (PE) parameters and performed as previously described (Chan et al., 2011 (link); Riley et al., 2014 (link)). All samples were analyzed with FloJo software (Tree Star).
Rabbit anti human c1q
Manufactured by Thermo Fisher Scientific
Rabbit anti-human C1q is a primary antibody that recognizes the C1q subcomponent of the first component of the human complement system. C1q is involved in the classical complement activation pathway and plays a crucial role in the immune response. This antibody can be used for the detection and analysis of C1q in various research and diagnostic applications.
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Lab products found in correlation
Alexa fluor 546 conjugated donkey anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Flojo software, by Tree Star (2 mentions)
Facscelesta cell analyzer, by BD (2 mentions)
Alexafluor 488 conjugated goat igg anti human igm, by Thermo Fisher Scientific (2 mentions)
Alexafluor 488 conjugated anti mouse igm clone af6 78, by Thermo Fisher Scientific (2 mentions)
2 protocols using rabbit anti human c1q
1
Rickettsia IgM and C1q Deposition Assay
2
Quantifying IgM and C1q Deposition on Rickettsia
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To assess IgM deposition, 1 × 107R. parkeri or R. australis bacilli were fixed in 4% PFA solution. After washing with PBS‐glycine, bacteria were incubated with 30% of NHS, IgM‐dplHS, or PBS for 1 hr at RT or overnight at 4°C. To assess both human IgM and C1q deposition on Rickettsia bacilli, all samples were treated with rabbit anti‐human C1q (Thermofisher), Alexa Fluor‐546‐conjugated donkey anti‐rabbit‐IgG (Invitrogen) and AlexaFluor‐488‐conjugated goat IgG anti‐human IgM (Invitrogen). Mouse IgM was measured as above but with C57BL6/J serum and AlexaFluor‐488‐conjugated anti‐mouse IgM (clone AF6‐78, Invitrogen). Fluorescence was assessed on a FACSCelesta Cell Analyzer (BD Biosciences) using fluorescein isothiocyanate (FITC) and phycoerythrin (PE) parameters and performed as previously described (Chan et al., 2011 (link); Riley et al., 2014 (link)). All samples were analyzed with FloJo software (Tree Star).
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