Anti-CD40L/CD40LG Antibody

Product Name

Anti-CD40L/CD40LG Antibody

View all CD40 Ligand/TNFSF5 Antibodies

SKU/Catalog Number

PA1020

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CD40L/CD40LG Antibody catalog # PA1020. Tested in IHC, WB applications. This antibody reacts with Human, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-CD40L/CD40LG Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1020)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of human CD40L, different from the related mouse sequence by three amino acids, rat sequence by four amino acids.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PA1020 is reactive to CD40LG in Human, Rat

Applications

PA1020 is guaranteed for IHC, WB Boster Guarantee

Observed Molecular Weight

29 kDa

Calculated molecular weight

29.274kDa

Background of CD40 Ligand/TNFSF5

CD40 ligand (CD40L) is a type II membrane protein of 261 amino acids on activated T cells that induces B cell proliferation and immunoglobulin secretion. It has homology with tumour necrosis factor-alpha and -beta, and has important functions in B-cell activation and differentiation. Human CD40L with 5 exons, is mapped to the proximal region of the mouse X chromosome on Xq26.3-27.1, and can be detected on T cells but is absent from B cells and monocytes. Since CD40L is expressed on platelets and released from them on activation, its predictive value as a marker for clinical outcome and the therapeutic effect of inhibition of glycoprotein IIb /IIIa receptor in patients with acute coronary syndromes was investigated. The soluble CD40L may be involved in the process of restenosis and that it exerts its effect by triggering a complex group of inflammatory reactions on endothelial and mononuclear cells. CD40L plays a central role in the pathophysiology of acute coronary syndromes, and has a role in the pathogenesis of coronary artery lesions.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, By Heat

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of sCD40L using anti-sCD40L antibody (PA1020).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: MCF-7 Whole Cell Lysate,
Lane 2: HELA Whole Cell Lysate,
Lane 3: JURKAT Whole Cell Lysate,
Lane 4: HMY2 Whole Cell Lysate,
Lane 5: COLO320 Whole Cell Lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-sCD40L antigen affinity purified polyclonal antibody (Catalog # PA1020) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for sCD40L at approximately 29KD. The expected band size for sCD40L is at 29KD.

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Figure 2. IHC analysis of sCD40L using anti-sCD40L antibody (PA1020).
sCD40L was detected in paraffin-embedded section of rat intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-sCD40L Antibody (PA1020) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of sCD40L using anti-sCD40L antibody (PA1020).
sCD40L was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-sCD40L Antibody (PA1020) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Gene/Protein Information For CD40LG (Source: Uniprot.org, NCBI)

Alternative Names

CD154 antigen; CD154; CD40 antigen ligand; CD40 Ligand; CD40L; CD40-L; CD40LG; CD40LIGM; gp39; hCD40L; HIGM1; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TNFSF5IMD3; TRAP; TRAPtumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome); tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand superfamily member 5 CD40LG CD154, CD40L, HIGM1, IGM, IMD3, T-BAM, TNFSF5, TRAP, gp39, hCD40L CD40 ligand CD40 ligand|CD40 antigen ligand|CD40-L|T-B cell-activating molecule|T-cell antigen Gp39|TNF-related activation protein|tumor necrosis factor (ligand) superfamily member 5

For more info on CD40LG, check out the CD40LG Infographic

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In this infographic, you will see the following information for CD40LG: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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