CD40 Antibody, anti-mouse

Name: CD40 Antibody, anti-mouse
Availability: InStock

CD40 Antibody, anti-mouseExtended Validation

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

Flow cytometry applications forCD40 Antibody, anti-mouse

APC
Biotin
FITC
PE
PE-Vio 770
Vio Bright FITC

Conjugate:

APC

Recommended dilution:

1:10

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-102-547

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:10)

Description:

Mouse spleen cells were stained with CD40 antibodies as well as with CD19-APC or CD19-PE and analyzed by flow cytometry. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD40 Antibody, anti-mouse

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:10

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-102-547

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Functional assay applications forCD40 Antibody, anti-mouse

pure-functional grade

Conjugate:

pure-functional grade

Products used:

130-093-022, 130-093-023

Type of functional assay:

blocking of receptor-ligand binding, in vitro and in vivo activation and maturation of antigen presenting cells

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 clones	Overlap 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.

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.

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.

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

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

Clone	FGK45.5
Clonality	monoclonal
Isotype	rat IgG2a
Isotype control	Isotype Control Antibody, rat IgG2a
Host	rat
Type of antibody	Primary antibodies, Functional-grade antibodies
Species	mouse
Antigen	CD40
Alternative names of antigen	Bp50, gp39, HIGM1, IGM, IMD3, T-BAM, TRAP, TNFRSF5, p50
Molecular mass of antigen [kDa]	30
Distribution of antigen	B cells, dendritic cells, endothelial cells, epithelial cells, fibroblasts, Langerhans cells, macrophages, mast cells, monocytes, platelets, lymphocytes, leukemia cells, plasma cells
Entrez Gene ID	21939
RRID	AB_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

Resources for CD40 Antibody, anti-mouse

Documents and Protocols

App notes/customer reports

Detection, enrichment, and direct phenotypic and functional characterization of antigen-specific naive and memory T cell subsets.

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD40 Antibody, anti-mouse

Publications

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
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

Applications

Extended validation