Clone:
FGK45.5
Type of antibody:
Primary antibodies, Functional-grade antibodies
Isotype:
rat IgG2a
Applications:
FA, FC, MICS, IF, IHC
Alternative names:
Bp50, gp39, HIGM1, IGM, IMD3, T-BAM, TRAP, TNFRSF5, p50

Extended validation for CD40 Antibody, anti-mouse

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 FGK45.5
3/23++
HM40-3++
65924+
1C10++
REA965++
Cells were incubated with an excess of purified unconjugated CD40 (FGK45.5) 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 CD40. C57BL6/6 splenocytes were stained with CD40 antibodies and with a suitable counterstaining. As a control, CD40 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 CD40. C57BL6/6 splenocytes were stained with CD40 antibodies and with a suitable counterstaining. As a control, CD40 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 CD40. C57BL6/6 splenocytes were stained with CD40 antibodies and with a suitable counterstaining. As a control, CD40 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 CD40. C57BL6/6 splenocytes were stained with CD40 antibodies and with a suitable counterstaining. As a control, CD40 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 CD40. C57BL6/6 splenocytes were stained with CD40 antibodies and with a suitable counterstaining. As a control, CD40 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 CD40. C57BL6/6 splenocytes were stained with CD40 antibodies and with a suitable counterstaining. As a control, CD40 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 CD40 (FGK45.5). 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 CD40 (FGK45.5). 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 CD40 (FGK45.5). 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 CD40 Antibody, anti-mouse

Overview

Clone FGK45.5 recognizes the mouse CD40 antigen, a 40–50 kDa glycoprotein and member of the tumor necrosis factor receptor (TNFR) family. CD40 is expressed on B lymphocytes, macrophages, and dendritic cells. It is an important costimulatory molecule for B cells as well as dendritic cells, monocytes, and other antigen-presenting cells (APCs). The interaction of CD40 on B cells with its ligand CD154 is involved in B cell activation and differentiation; engagement of CD40 on dendritic cells leads to maturation.
Clone FGK45 can be used for blocking of CD40/CD154 interaction and for
in vitro
and
in vivo
activation of CD40 expressing APCs.

Alternative names

Bp50, gp39, HIGM1, IGM, IMD3, T-BAM, TRAP, TNFRSF5, p50

Detailed product information

Technical specifications

CloneFGK45.5
Clonalitymonoclonal
Isotyperat IgG2a
Isotype controlIsotype Control Antibody, rat IgG2a
Hostrat
Type of antibodyPrimary antibodies, Functional-grade antibodies
Speciesmouse
AntigenCD40
Alternative names of antigenBp50, gp39, HIGM1, IGM, IMD3, T-BAM, TRAP, TNFRSF5, p50
Molecular mass of antigen [kDa]30
Distribution of antigenB cells, dendritic cells, endothelial cells, epithelial cells, fibroblasts, Langerhans cells, macrophages, mast cells, monocytes, platelets, lymphocytes, leukemia cells, plasma cells
Entrez Gene ID21939
RRIDAB_871725, AB_871724, AB_2660756, AB_2660757, AB_2660758, AB_2660759, AB_2660760, AB_2660761, AB_2660762, AB_2660763, AB_2660764, AB_2660765, AB_2660766

References for CD40 Antibody, anti-mouse

Publications

  1. Rolink, A. et al. (1996) The SCID but not the RAG-2 gene product is required forS mu-S epsilon heavy chain class switching. Immunity 5(4): 319-330
  2. Bonifaz, L. et al. (2002)
    Efficient targeting of protein antigen to the dendritic cell receptor DEC-205 in the steady state leads to antigen presentation on major histocompatibility complex class I products and peripheral CD8
    +
    T cell tolerance.
    J. Exp. Med. 196: 1627-1638

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