46-1548-42

antibody from Invitrogen Antibodies

Targeting: CD40LG CD154, CD40L, gp39, hCD40L, HIGM1, IMD3, TNFSF5, TRAP

Provider product page for 46-1548-42

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [4]
Comments [0]
Validations
Flow cytometry [1]
Other assay [3]

Submit

Product number: 46-1548-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD154 (CD40 Ligand) Monoclonal Antibody (24-31), PerCP-eFluor™ 710, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The 24-31 monoclonal antibody reacts with human CD154, a 39 kDa transmembrane glycoprotein also known as gp39 and CD40 ligand (CD40L). CD154 is a member of the TNF superfamily and is expressed transiently by activated T cells. Through its binding to CD40 on antigen presenting cells including B cells, monocytes/macrophages and dendritic cells, CD154 serves a crucial function in T-APC cognate interaction. CD154 interaction with CD40 transduces signals for T-dependent B-cell activation and induces B cell cycle entry. 24-31 cross-reacts with the cynomolgus monkey CD154. Applications Reported: This 24-31 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 24-31 antibody has been pre-titrated and tested by flow cytometric analysis of stimulated normal human peripgheral blood cells. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. PerCP-eFluor® 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm). Please make sure that your instrument is capable of detecting this fluorochrome. For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative. Our testing indicates that PerCP-eFluor® 710 conjugated antibodies are stable when stained samples are exposed to freshly prepared 2% formaldehyde overnight at 4°C, but please evaluate for alternative fixation protocols. Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 24-31
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

CD40LG protein structure - 46-1548-42 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 SARS-CoV-2 Infection Induces Durable, Functional Spike-Reactive CD4 + T Cells (A) Representative flow cytometry plots of ICOS and CD40L expression on antigen-experienced (non-CD45RA + CCR7 + ) CD4 + T cells 20 h after incubation of PBMCs from HC and CoV2 + individuals at V1 and V2 with vehicle or SARS-CoV-2 spike. (B) Number of antigen-experienced ICOS + CD40L + CD4 + T cells per 1x10 6 CD4 + T cells from HC and CoV2 + samples after incubation with vehicle (Veh.) or spike (S) at both time points (right) and calculated number of spike-responsive CD4 + T cells (number after incubation with spike minus number after incubation with vehicle) compared across time points (left). (C) Number of antigen-experienced CXCR5 + ICOS + CD40L + CD4 + T cells (cTfh) per 1 x 10 6 CD4 + T cells from HC and CoV2 + samples after incubation with Veh. or S at both time points (right) and calculated number of spike-responsive cTfh cells (number after incubation with spike minus number after incubation with vehicle) compared across time points (left). (D) Representative flow cytometry plots of sorted CD4 + naive (CD45RA + CCR7 + ), T CM (CD45RA - CCR7 + ), or T EM (CD45RA - CCR7 - ) T cells from HC and CoV2 + PBMCs after 5-6 days of co-culture with SARS-CoV-2 spike-protein-pulsed autologous monocytes and measuring proliferation by CPD dilution. (E) SARS-CoV-2 spike-specific expansion of sorted CD4 + naive T, T CM , and T EM cells from V1 (circles) and V2 (squares) reported as frequency of CXC

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. The remnants of LM8 decreased the co-stimulatory molecules, and inhibited IL-12 and IL-6 levels, increased IL-10 levels, and inhibited the ability of DCs to stimulate T-cell proliferation. (A) The DCs were isolated following a 48-h co-culture with the remnants of LM8 cells. The cells were then labeled with antibodies against CD11c, MHC-2, CD40, CD86, CD80 and CCR7 for phenotypic analysis by flow cytometry. The numbers in the histograms indicate the geometric mean fluorescence intensity. (B) Following isolation from the co-culture system, the DCs were cultured for 12 h. Subsequently, the expression levels of IL-12, IL-6 and IL-10 in the supernatant were analyzed by ELISA. (C) CD4 + T cells from DO11.10 OVA 323-339 -specific (TCR-transgenic x C57BL/6) F1 hybrid mice were co-cultured with DCs or mDCs (control) in the presence of OVA peptides. Five days later, the total number of viable CD4 + T (CD4 + 7-AAD - ) cells in each well was measured by flow cytometry. Data represent one of at least three experiments with similar results. *P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. PDT treatment partly reversed the effect of LM8 remnants on the phenotype of DCs and their ability to stimulate T cells proliferation. (A) The LM8 cells were pre-treated with PDT, and then labeled with antibodies against MHC-2, CD11c, CD40, CD86, CD80 and CCR7, for phenotypic analysis by flow cytometry. (B) The numbers in the histograms indicate the geometric mean fluorescence intensity. *P