Clone:
FN50
Type of antibody:
Primary antibodies
Isotype:
mouse IgG1κ
Applications:
FC, ICFC, MICS, IF, IHC, MC
Alternative names:
AIM, BL-AC/P26, CLEC2C, EA1, GP32/28, MLR-3, VEA

Extended validation for CD69 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 FN50
REA824++
L78++
Cells were incubated with an excess of purified unconjugated CD69 (FN50) 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 CD69. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with CD69 antibodies and with a suitable counterstaining. As a control, CD69 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) 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 CD69. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with CD69 antibodies and with a suitable counterstaining. As a control, CD69 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) 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 CD69. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with CD69 antibodies and with a suitable counterstaining. As a control, CD69 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) 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 CD69. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with CD69 antibodies and with a suitable counterstaining. As a control, CD69 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) 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 CD69. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with CD69 antibodies and with a suitable counterstaining. As a control, CD69 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) 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 CD69. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with CD69 antibodies and with a suitable counterstaining. As a control, CD69 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD69 (FN50). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD69 (FN50). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD69 (FN50). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD69 Antibody, anti-human

Overview

Clone FN50 recognizes the human CD69 antigen, also known as activation inducer molecule (AIM). CD69 is involved in the early events of lymphocyte, monocyte, and platelet activation. Cross-linking of CD69 induces cytotoxic activity and costimulates cytokine production of activated NK cells and selected T cell clones. CD69 is transiently expressed on activated leukocytes including T cells, thymocytes, B cells, NK cells, neutrophils, and eosinophils. It is constitutively expressed by a subset of medullary mature thymocytes, platelets, mantle B cells, and certain CD4
+
T cells in germinal centers of normal lymph nodes.

Alternative names

AIM, BL-AC/P26, CLEC2C, EA1, GP32/28, MLR-3, VEA

Detailed product information

Technical specifications

CloneFN50
Clonalitymonoclonal
Isotypemouse IgG1κ
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
, baboon
AntigenCD69
Alternative names of antigenAIM, BL-AC/P26, CLEC2C, EA1, GP32/28, MLR-3, VEA
Molecular mass of antigen [kDa]23
Distribution of antigenB cells, Langerhans cells, leukocytes, lymphocytes, NK cells, platelets, T cells, eosinophils, neutrophils, thymocytes
Entrez Gene ID969
RRIDAB_2733656, AB_2751192, AB_2751126, AB_2733401, AB_2733402, AB_2733950, AB_2733951, AB_2784272, AB_2784271, AB_2733139, AB_2733140, AB_2751191, AB_2751125, AB_2661139, AB_2733655

References for CD69 Antibody, anti-human

Publications

  1. Lin, G. X. et al. (2002) Cross-reactivity of CD antibodies in eight animal species. In: Leucocyte typing VII, Oxford University Press. : 519-524

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