biotin conjugated anti human kappa light chain antibody  (Absolute Biotech Inc)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human FcRL4 and FcRL5 are receptors for IgA and IgG 1 , 2

doi: 10.4049/jimmunol.1102651

Figure Lengend Snippet: HEK293T cells were transiently transfected with human cDNA for FcRL1, 3, 4, 5, 6 and CD200R expressed in pFLAG-CMV-3, or CD32 in pEF6. Binding of heat-aggregated human serum IgG and IgA (Y-axis) to cells expressing human FcRL4 and FcRL5 was easily observable by FACS. Cell surface expression of each protein was confirmed using FITC-conjugated anti-FLAG M2 monoclonal antibody (X-axis), or by staining with anti-CD32 antibody. Binding to CD200R and CD32 are included as negative and positive controls respectively. No binding of human IgM to any of the FcRL proteins was observed (not shown).

Article Snippet: 293 cells were incubated for 30 minutes on ice with the Igs and washed four times, followed by incubation with biotin-conjugated goat F(ab’)2 anti-human kappa light chain antibody (for IgG1, IgG2, IgG3 and IgG4), or a combination of biotinylated goat F(ab’)2 anti-human kappa and goat F(ab’)2 anti-human lambda antibody (for total IgG and IgA) (Southern Biotech) for 20 minutes on ice.

Techniques: Transfection, Binding Assay, Expressing, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human FcRL4 and FcRL5 are receptors for IgA and IgG 1 , 2

doi: 10.4049/jimmunol.1102651

Figure Lengend Snippet: A) Human IgG isotypes were assayed by FACS for binding to FcRL4, FcRL5, and CD32. Dark lines indicate aggregated IgG bound to the indicated receptor-transfected 293 cells. Gray shading indicates binding of Igs to negative control CD200R-transfected cells. Histograms show populations gated on receptor-expressing cells by simultaneous staining with anti-Flag (FcRL4, FcRL5 and CD200R) or anti-CD32 antibodies B) Portions of the ectodomain of FcRL5 comprising Ig domains 1-3, 4-6, or 7-9 were expressed on the surface of 293 cells. Binding of IgG1 mock ICs to FcRL5 d1-3 was measured by FACS.

Article Snippet: 293 cells were incubated for 30 minutes on ice with the Igs and washed four times, followed by incubation with biotin-conjugated goat F(ab’)2 anti-human kappa light chain antibody (for IgG1, IgG2, IgG3 and IgG4), or a combination of biotinylated goat F(ab’)2 anti-human kappa and goat F(ab’)2 anti-human lambda antibody (for total IgG and IgA) (Southern Biotech) for 20 minutes on ice.

Techniques: Binding Assay, Transfection, Negative Control, Expressing, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human FcRL4 and FcRL5 are receptors for IgA and IgG 1 , 2

doi: 10.4049/jimmunol.1102651

Figure Lengend Snippet: A) FACS staining of FcRL5 d1-3 or d4-6 with Mab 501H9 and 509F6. B) Pre-incubation of antibody 509F6 with FcRL5-expressing 293 cells prevents binding of FcRL5-transfected 293 cells to mock ICs. C) Blockade of IgA binding to FcRL4 by anti-FcRL4 clone 413D12 (IgG2b) but not isotype matched anti-FcRL4 clone 418C8. D) Staining of tonsillar B cells with anti-FcRL4 (413D12) and anti-FcRL5 (509F6) monoclonal antibodies or isotype controls. (Plots show cells following gating on live CD19+ cells.) E) Anti-FcRL4 and anti-FcRL5 identify mixed populations of CD27+ and CD27− tonsillar B cells. F) All FcRL4+ tonsillar B cells co-express FcRL5. E,F) Plots are representative of results obtained with tonsil samples from five different donors.

Article Snippet: 293 cells were incubated for 30 minutes on ice with the Igs and washed four times, followed by incubation with biotin-conjugated goat F(ab’)2 anti-human kappa light chain antibody (for IgG1, IgG2, IgG3 and IgG4), or a combination of biotinylated goat F(ab’)2 anti-human kappa and goat F(ab’)2 anti-human lambda antibody (for total IgG and IgA) (Southern Biotech) for 20 minutes on ice.

Techniques: Staining, Incubation, Expressing, Binding Assay, Transfection