Clone:
3A3-N2
Type of antibody:
Primary antibodies
Isotype:
mouse IgG1κ
Applications:
ICFC
Alternative names:
CVID11, Za11

Extended validation for IL-21 Antibody, anti-human

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with 3A3-N2
4BG1-
REA1039++
2B2-G20-
Cells were incubated with an excess of purified unconjugated IL-21 (3A3-N2) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for IL-21. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-21 antibodies. As a control, IL-21 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used
View details
Flow cytometric comparison of different clones for IL-21. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-21 antibodies. As a control, IL-21 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used
View details
Flow cytometric comparison of different clones for IL-21. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-21 antibodies. As a control, IL-21 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used
View details
Flow cytometric comparison of different clones for IL-21. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-21 antibodies. As a control, IL-21 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used
Flow cytometric comparison of different clones for IL-21. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-21 antibodies. As a control, IL-21 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used

Specifications for IL-21 Antibody, anti-human

Overview

Interleukin 21 (IL-21) is secreted by a certain subset of CD4
+
T cells, called T follicular helper (Tfh) cells, also characterized by the expression of CXCR5 and ICOS and the lineage-specific transcription factor BCL6. But IL-21 production can also be found in other T helper (Tʜ) cell lineages and natural killer T (NKT) cells. IL-21 acts in a autocrine manner on Tfh cells and is critical for antibody production by B cells, it enhances natural killer (NK) cell functions, and promotes proliferation of CD8
+
T cells.

Alternative names

CVID11, Za11

Detailed product information

Technical specifications

Clone3A3-N2
Clonalitymonoclonal
Isotypemouse IgG1κ
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman
AntigenIL-21
Alternative names of antigenCVID11, Za11
Molecular mass of antigen [kDa]15
Entrez Gene ID59067
RRIDAB_2801779, AB_2659997, AB_2659996, AB_2801784

Resources for IL-21 Antibody, anti-human

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

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