Anti-CD40L antibody (ab2391)

Overview

  • Product name
  • Description
    Rabbit polyclonal to CD40L
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, ELISA, WB, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide surrounding amino acid 246 of human CD40 Ligand

  • Positive control
    • Human muscle tissue sections

Properties

Applications

Our Abpromise guarantee covers the use of ab2391 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/200.
ICC/IF 1/200. See Abreview
ELISA Use a concentration of 0.5 - 1 µg/ml.
WB Use a concentration of 1 µg/ml.
IHC-P Use at an assay dependent concentration.
IHC-Fr 1/100. Fix with acetone -20°C for 10 min (provides a better signal than unfixed tissue; PFA 4% abolishes the signal). Incubate primary for 24h at 4°C. Use amplification (indirect IF).

Target

  • Function
    Mediates B-cell proliferation in the absence of co-stimulus as well as IgE production in the presence of IL-4. Involved in immunoglobulin class switching.
    Release of soluble CD40L from platelets is partially regulated by GP IIb/IIIa, actin polymerization, and an matrix metalloproteinases (MMP) inhibitor-sensitive pathway.
  • Tissue specificity
    Specifically expressed on activated CD4+ T-lymphocytes.
  • Involvement in disease
    Defects in CD40LG are the cause of X-linked immunodeficiency with hyper-IgM type 1 (HIGM1) [MIM:308230]; also known as X-linked hyper IgM syndrome (XHIM). HIGM1 is an immunoglobulin isotype switch defect characterized by elevated concentrations of serum IgM and decreased amounts of all other isotypes. Affected males present at an early age (usually within the first year of life) recurrent bacterial and opportunistic infections, including Pneumocystis carinii pneumonia and intractable diarrhea due to cryptosporidium infection. Despite substitution treatment with intravenous immunoglobulin, the overall prognosis is rather poor, with a death rate of about 10% before adolescence.
  • Sequence similarities
    Belongs to the tumor necrosis factor family.
  • Post-translational
    modifications
    The soluble form derives from the membrane form by proteolytic processing.
    N-linked glycan is a mixture of high mannose and complex type. Glycan structure does not influence binding affinity to CD40.
    Not O-glycosylated.
  • Cellular localization
    Secreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD 40L antibody
    • CD154 antibody
    • CD40 antigen ligand antibody
    • CD40 ligand antibody
    • CD40 ligand, soluble form antibody
    • CD40-L antibody
    • CD40L antibody
    • CD40L_HUMAN antibody
    • CD40LG antibody
    • gp39 antibody
    • hCD40L antibody
    • HIGM1 antibody
    • IGM antibody
    • IMD3 antibody
    • T B cell activating molecule antibody
    • T BAM antibody
    • T-cell antigen Gp39 antibody
    • TNF-related activation protein antibody
    • TNFSF5 antibody
    • TrAP antibody
    • Tumor necrosis factor (ligand) superfamily member 5 antibody
    • Tumor necrosis factor ligand superfamily member 5 antibody
    see all

Images

  • ab2391 staining CD40L in Human platelet cells by Flow cytometry.
    Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton-X-100 in 2% BSA for 15 minutes. Primary antibody used at a 1/200 dilution and incubated for 16 hours at 4°C. The secondary antibody used was an Alexa Fluor®488 conjugated chicken anti-rabbit IgG (H+L) at a 1/500 dilution.

    See Abreview

  • Immunofluorescent analysis of paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells CD40L with ab2391 at 1/200 dilution, followed by Chicken Anti-Rabbit IgG H&L (Alexa Fluor® 488 secondary antibody at 1/750 dilution (green). WGA is detected with Alexa Fluor® 568 Wheat Germ Agglutinin (red). The nuclear counter stain is DAPI (blue). 

  • ab2391 at a 1/100 dilution staining CD40L from human inflamed muscle tissue sections by immunohistochemistry (frozen sections). The tissue was acetone fixed (this provides a better result than unfixed tissue) and then incubated with the antibody for 24 hours at 4 °C. Alexa-Fluor ® 594 was used as the secondary antibody.

    See Abreview

  • ab2391 at 1/500 staining rat spleen tissue sections by IHC-P. The tissue was fomraldehyde fixed and blocked with serum prior to incubation with the antibody for 18 hours. An Alexa-Fluor ® 488 conjugated goat anti-rabbit was used as the secondary.

    See Abreview

References

ab2391 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Te agradezco mucho que te hayas tomado la molestia de enviarnos las imágenes y el protocolo.
Efectivamente, se observa una tinción inespecífica en los tejidos utilizados, y probablemente haya habido algún problema con la alícuota de anticuerpo rec...

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Cualquier comentario o queja procedente de nuestros clientes es muy importante para mejorar y monitorizar la calidad de nuestros productos.
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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RAW 264.7)
Specification
RAW 264.7
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 15min
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted May 23 2012

Application
Immunocytochemistry
Sample
Human Cell (Monocytes from Apheresis)
Specification
Monocytes from Apheresis
Fixative
BD Cytofix/Cytoperm 20 min
Permeabilization
Yes - BD Cytofix/Cytoperm 20 min
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 2°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Feb 02 2012

Thank you for contacting Abcam regarding our CD40 and CD40L antibodies. Below please find the requested information: ab2391 Rabbit Polyclonal to CD40L - Unfortunately the specific immunogen sequence is proprietary, however I can inform you that...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Platelets)
Loading amount
20 µg
Specification
Platelets
Treatment
A23187 for 1 h
Gel Running Conditions
Reduced Denaturing (4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Nov 14 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Platelets)
Specification
Platelets
Preparation
Cell harvesting/tissue preparation method: PL were isolated spinning Platelet rich plasma on Histopaque
Sample buffer: PBS
Fixation
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 15min
Gating Strategy
Platelets
Username

Abcam user community

Verified customer

Submitted Dec 31 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (spinal cord homogenate)
Loading amount
20 µg
Specification
spinal cord homogenate
Treatment
sham or nerve transected
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Aug 06 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (spleen)
Specification
spleen
Fixative
Formaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Ms. Nancy Nutile-McMenemy

Verified customer

Submitted Jul 11 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (spleen)
Specification
spleen
Fixative
Formaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Ms. Nancy Nutile-McMenemy

Verified customer

Submitted Jul 06 2007

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