Mouse Anti-Migis-α Recombinant Antibody (clone 29C11) (CAT#: FAMAB-0227CQ)

Figure 1 A graph which depicts an ELISA showing the reactivity of an anti-IgA.

Fc mAb (3C10) and two anti-migis-α mAbs (8G7 and 29C11) with human IgA and constructs, which contain various human membrane-bound α chain, or other components.

Figure 2 Epitope mapping of anti-migis-α mAbs in ELISA.

A shows binding reactivity of anti-migis-α mAbs with different parts of migis-αL. B shows binding reactivity of anti-migis-α mAbs with short peptides of the N-terminal part of migis-αL.

Figure 3 Determination of the relative affinity of 8G7 and 29C11 in binding to ma in ELISA.

A shows binding strength of 8G7 and 29C11 against mα1.FcL-456S-LZ proteins. B. shows relative ability of 8G7 and 29C11 in competing with 200 nM biotin-conjugated 29C11 in binding to FcL-456S-LZ protein.

Figure 4 Staining profiles of anti-migis-α mAbs on DAKIKI cells and mα1.Fc-expressing Ramos transfectomas.

The gray profiles are negative control mAbs of the same IgG isotypes.

Figure 5 The reactivity of anti-migis-α mAbs toward DAKIKI cells with or without MβCD treatments.

Figure 6 Results indicating that 3C10 and 8G7 can induce apoptosis in two mα1.Fc-expressing Ramos transfectomas, whereas 29C11 fails to do so.

Figure 7 Recombinant proteins used in this study and the reactivity of anti-migis-α mAbs with these proteins as detected by ELISA

Reactivity of anti-migis-α mAbs with Proteins I, II, and III as detected by ELISA. The mAbs 7C3, 9F5, and 29C11 were developed in the present study. 3C10, an anti-IgA1.Fc mAb; 4B12 (IgG1κ) and a20 (IgG2aκ), both specific for "CεmX", a domain located between CH4 and the migis segment of mε, used as isotype controls; mεFcL-LZ, a recombinant protein of mε.

Hung, A. F. H., Chen, J. B., Lu, C. S., Chen, N. Y., Yu, H. M., & Chang, T. W. (2011). Lipid rafts hinder binding of antibodies to the extracellular segment of the membrane-anchor peptide of mIgA. Molecular immunology, 48(15-16), 1975-1982.

Figure 8 Competition of mAbs 7C3 and 9F5 with 29C11 for binding to migis-α.

Biotin-conjugated 29C11 was mixed with various concentrations of mAbs before the mixtures were placed in microtiter wells coated with mFcL-LZ. Following incubation, HRP-conjugated avidin and its substrate, TMB, were applied for colorimetric measurements.

Hung, A. F. H., Chen, J. B., Lu, C. S., Chen, N. Y., Yu, H. M., & Chang, T. W. (2011). Lipid rafts hinder binding of antibodies to the extracellular segment of the membrane-anchor peptide of mIgA. Molecular immunology, 48(15-16), 1975-1982.

Figure 9 Ability of anti-migisα mAbs to capture mIgA from lysed B cells.

DAKIKI cells and Daudi transfectants lysed in ice-cold detergent-containing buffer were incubated with a mAb and Protein G beads were added to absorb the mAb and its complex. Pulled mixture was then subjected to Western blot analysis. The proteins in the mixture were resolved in polyacrylamide gel electrophoresis (10% SDS), transferred to PVDF membrane, reacted with HRP-conjugated goat anti-human IgA or anti-mouse IgG, and analyzed by chemo-luminescence. (A) Cells lysed in TNE buffer containing 1% Triton X-100; (B) Cells lysed in RIPA buffer; (C) Cells treated with 10 mM MCD and then lysed in TNE buffer containing 1% Triton X-100.

Hung, A. F. H., Chen, J. B., Lu, C. S., Chen, N. Y., Yu, H. M., & Chang, T. W. (2011). Lipid rafts hinder binding of antibodies to the extracellular segment of the membrane-anchor peptide of mIgA. Molecular immunology, 48(15-16), 1975-1982.

Figure 10 Effects of mIgA cross-linking on the staining of mIgA-expressing B cells by 29C11.

(A) The binding of mAb 3C10, but not 29C11,to mFcL-LZ is inhibited by goat IgG anti-human IgA (anti-huIgA) as revealed by ELISA. In this assay, non-immunized goat IgG or goat anti-human IgA were incubated with mFcL-LZ, before 3C10 or 29C11 was added. HRP-conjugated goat anti-mouse IgG was used for colorimetric measurements. (B) DAKIKI and Daudi/mFcL cells were incubated with goat IgG or goat anti-human IgA and then stained by 3C10 or 29C11. FITC-conjugated goat F(ab)2 anti-mouse IgG were used for flow cytometric analysis. Histograms–shaded, cells incubated without 3C10 or 29C11; grey lines, cells incubated with goat IgG; black lines, cells incubated with goat IgG anti-human IgA.

Hung, A. F. H., Chen, J. B., Lu, C. S., Chen, N. Y., Yu, H. M., & Chang, T. W. (2011). Lipid rafts hinder binding of antibodies to the extracellular segment of the membrane-anchor peptide of mIgA. Molecular immunology, 48(15-16), 1975-1982.