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Anti igκ

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Anti-Igκ is a laboratory reagent used in immunochemical assays to detect and quantify the kappa light chain (Igκ) of immunoglobulins. It functions as a specific binding agent for the Igκ antigen.

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Lab products found in correlation

Facscalibur, by BD (2 mentions) Cellquest, by BD (2 mentions) Anti cd25 pe, by BD (2 mentions) Anti cd4 fitc, by BD (2 mentions) Anti cd69 pe, by BD (2 mentions) Anti cd5 pe, by BD (2 mentions) Anti cd43 pe, by BD (2 mentions) Anti b220 fitc, by BD (2 mentions) Anti b220 percp, by BD (2 mentions) Pe anti igκ, by BD (2 mentions) Pe anti cd138, by BD (2 mentions) Anti cd86 pe, by BD (2 mentions) Anti cd80 pe, by BD (2 mentions) Anti igm pe, by BD (2 mentions) Anti mouse ceacam1 mab, by BD (2 mentions) Pe anti h 2kd, by BD (2 mentions) Pe anti 1 ad, by BD (2 mentions) Fluo 4, by Dojindo Laboratories (2 mentions) A20 cell line, by American Type Culture Collection (2 mentions) Pe anti cd3ε, by BD (2 mentions)

Spelling variants of this product

Goat F(ab’)2 anti-mouse Igκ (8 citations) HRP-labeled goat anti-human Igκ antibody (6 citations) Goat anti-mouse Igκ (4 citations) Anti-mouse Igκ and Igλ antibodies (3 citations) Anti-human Igκ (2 citations) Anti-human-Igκ-FITC (2 citations) Goat anti-mouse Igκ and anti-mouse Igλ polyclonal antibodies (2 citations) Goat anti-mouse Igκ-HRP (2 citations)

4 protocols using anti igκ

1

Isolation and Analysis of Insulin-Binding B Cells

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Splenocytes were treated with 2.4G2 anti-Fc γRIIB antibody (Cambier lab) to block FcR binding. In primary staining, cells were incubated with anti-B220 (BD, RA3-6B2) and anti-IgM antibodies (Cambier lab, b-7-6), and 0.1μg biotinylated human recombinant insulin (Sigma) for 30min on ice. In secondary staining, cells were stained with 2µg Alexa Fluor™ 647 conjugated streptavidin (Invitrogen) for 20min on ice. Cells were washed and resuspended in MACS buffer (PBS/0.5% BSA/2 mmol/L EDTA), and incubated with anti-Cy5/anti-Alexa 647 microbeads (Miltenyi Biotec) for 15min at 4°C. To enrich IBCs, cells were passed through LS column mounted on a magnet (Miltenyi Biotec). To distinguish kappa (Igκ) and lambda (Igλ) light chains, cell markers such as: anti-Igκ (SouthernBiotech) and anti-Igλ (SouthernBiotech) were used during primary staining. Flow cytometry and cell sorting were performed using an LSR Fortessa X-20 (BD) and FACS on MoFlo Astrios EQ (Beckman Coulter). Data were analyzed with FlowJo software version 9 (Ashland, OR, USA).
Banach M., Harley I.T., Getahun A, & Cambier J.C. (2022). Comparative analysis of the repertoire of insulin-reactive B cells in type 1 diabetes-prone and resistant mice. Frontiers in Immunology, 13, 961209.
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2

Characterization of A20 B cell lymphoma

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A20 cell line, derived from mouse B cell lymphoma on BALB/c background [13 (link)], was obtained from American Type Culture Collection (Manassas, VA). Primary cells were isolated from bone marrow (BM), spleen (SPL) or peripheral blood (PB) of B6 by standard methods. The A20 transfectants or primary cells were stained with the following antibodies (Abs): anti-mouse CEACAM1 mAb, CC1 (kindly provided by Dr. Kathlyn Holmes, University of Colorado, CO), anti-IgG1, FITC-anti-B220, PE-anti-IgM, FITC-anti-CD4, PE-anti-CD3ε, PerCP-anti-B220, PE-anti-CD43, PE-anti-CD25, PE-anti-Igκ, PE-anti-CD138, PE-anti-CD5, PE-anti-H-2Kd, PE-anti-I-Ad, PE-anti-CD69, PE-anti-CD80 and PE-anti-CD86 Abs (BD Biosciences, San Jose, CA). Data acquisition was performed using FACS Calibur™ and analysis software Cell Quest™ (BD Biosciences).
In some experiments, the A20 transfectants were treated with Fluo-4® (Dojindo, Kumamoto, Japan), and applied for FACS to analyze intracellular Ca2+ influx. During the acquisition, cells were treated with anti-Igκ (Southern Biotech, Birmingham, AL) to stimulate BCR signaling, and kinetics for Ca2+ influx were measured at FL2.
Tsugawa N., Yamada D., Watabe T., Onizawa M., Wang S., Nemoto Y., Oshima S., Tsubata T., Adachi T., Kawano Y., Watanabe M., Blumberg R.S., Okamoto R, & Nagaishi T. (2020). CEACAM1 specifically suppresses B cell receptor signaling-mediated activation. Biochemical and biophysical research communications, 535, 99-105.
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3

Characterization of A20 B cell lymphoma

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A20 cell line, derived from mouse B cell lymphoma on BALB/c background [13 (link)], was obtained from American Type Culture Collection (Manassas, VA). Primary cells were isolated from bone marrow (BM), spleen (SPL) or peripheral blood (PB) of B6 by standard methods. The A20 transfectants or primary cells were stained with the following antibodies (Abs): anti-mouse CEACAM1 mAb, CC1 (kindly provided by Dr. Kathlyn Holmes, University of Colorado, CO), anti-IgG1, FITC-anti-B220, PE-anti-IgM, FITC-anti-CD4, PE-anti-CD3ε, PerCP-anti-B220, PE-anti-CD43, PE-anti-CD25, PE-anti-Igκ, PE-anti-CD138, PE-anti-CD5, PE-anti-H-2Kd, PE-anti-I-Ad, PE-anti-CD69, PE-anti-CD80 and PE-anti-CD86 Abs (BD Biosciences, San Jose, CA). Data acquisition was performed using FACS Calibur™ and analysis software Cell Quest™ (BD Biosciences).
In some experiments, the A20 transfectants were treated with Fluo-4® (Dojindo, Kumamoto, Japan), and applied for FACS to analyze intracellular Ca2+ influx. During the acquisition, cells were treated with anti-Igκ (Southern Biotech, Birmingham, AL) to stimulate BCR signaling, and kinetics for Ca2+ influx were measured at FL2.
Tsugawa N., Yamada D., Watabe T., Onizawa M., Wang S., Nemoto Y., Oshima S., Tsubata T., Adachi T., Kawano Y., Watanabe M., Blumberg R.S., Okamoto R, & Nagaishi T. (2020). CEACAM1 specifically suppresses B cell receptor signaling-mediated activation. Biochemical and biophysical research communications, 535, 99-105.
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4

Quantification of Anti-DNA Antibodies

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Total splenic B cells, marginal zone B cells, transitional B cells, or
Igκ-depleted B cells were cultured at 106 cells/ml in complete
RPMI media with or without 5 mg/ml LPS (Sigma). Supernatants were harvested
after 5 days of culture.
Anti-dsDNA and/or anti-ssDNA ELISAs were performed on serial dilutions
of serum (1:100, 1:400, 1:1600) or culture supernatant (neat, 1:5, 1:25, 1:125)
as described in (25 (link)). Anti-IgM, IgG,
anti-Igκ, or anti-Igλ alkaline phosphatase-conjugated detection
antibodies (Southern Biotech) were used as indicated in the figure legends.
Ottens K., Hinman R.M., Barrios E., Skaug B., Davis L.S., Li Q.Z., Castrillon D.H, & Satterthwaite A.B. (2018). Foxo3 promotes apoptosis of BCR-stimulated immature B cells, thus limiting the window for receptor editing. Journal of immunology (Baltimore, Md. : 1950), 201(3), 940-949.
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