CD40L Recombinant Rabbit Monoclonal Antibody [JM11-34]
cat.: ET1703-64
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JM11-34
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 29 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human CD40L aa 103-150 / 261.
Positive control: Jurkat cell lysate, K-562 cell lysate, PC-12 cell lysate, mouse liver tissue lysate, rat thymus tissue lysate, Hela, MCF-7, NIH/3T3, human spleen tissue, THP-1.
Subcellular location: Cell membrane, Cell surface, Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:5,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P29965 Human | P27548 Mouse | Q9Z2V2 Rat
Alternative names: CD 40L CD154 CD40 antigen ligand CD40 ligand CD40 ligand, soluble form CD40-L CD40L CD40L_HUMAN CD40LG gp39 hCD40L HIGM1 IGM IMD3 T B cell activating molecule T BAM T-cell antigen Gp39 TNF-related activation protein TNFSF5 TrAP Tumor necrosis factor (ligand) superfamily member 5 Tumor necrosis factor ligand superfamily member 5
Images
ET1703-64_1.jpg Fig1: Western blot analysis of CD40L on different lysates with Rabbit anti-CD40L antibody (ET1703-64) at 1/5,000 dilution.

Lane 1: Jurkat cell lysate (15 µg/Lane)
Lane 2: K-562 cell lysate (15 µg/Lane)
Lane 3: PC-12 cell lysate (15 µg/Lane)
Lane 4: Mouse liver tissue lysate (20 µg/Lane)
Lane 5: Rat thymus tissue lysate (20 µg/Lane)

Predicted band size: 29 kDa
Observed band size: 40 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-64) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1703-64_2.jpg Fig2: ICC staining of CD40L in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-64, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-64_3.jpg Fig3: ICC staining of CD40L in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-64, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-64_4.jpg Fig4: ICC staining of CD40L in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-64, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-64_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD40L antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-64, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-64_6.jpg Fig6: Flow cytometric analysis of CD40L was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-64, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.