CD40 Mouse Monoclonal Antibody [A1E6]
cat.: EM1901-40
Product Type: Mouse monoclonal IgG2b, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: A1E6
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 31 kDa
Isotype: IgG2b
Immunogen: Recombinant protein within human CD40 aa 1-250.
Positive control: Raji cell lysate, Daudi cell lysate, MG-63, SiHa, SKOV-3, human tonsil tissue.
Subcellular location: Cell membrane, secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P

1:500-1:2,000
1:50-1:100
1:1,000
Uniprot #: SwissProt: P25942 Human
Alternative names: AI326936 B cell associated molecule CD40 B cell surface antigen CD40 B cell-associated molecule B-cell surface antigen CD40 Bp50 CD 40 CD40 CD40 antigen (TNF receptor superfamily member 5) CD40 antigen CD40 molecule CD40 molecule, TNF receptor superfamily member 5 CD40 protein CD40 type II isoform CD40L receptor CDw40 GP39 HIGM1 IGM IMD3 MGC9013 Nerve growth factor receptor related B lymphocyte activation molecule OTTHUMP00000031699 OTTHUMP00000031700 p50 T-BAM TBAM TNF receptor superfamily member 5 TNFRSF5 TNR5_HUMAN TRAP Tumor necrosis factor receptor superfamily , member 5 Tumor necrosis factor receptor superfamily member 5 Tumor necrosis factor receptor superfamily member 5 precursor Tumor necrosis factor receptor superfamily, member 5, isoform CRA_a
Images
EM1901-40_1.jpg Fig1: Western blot analysis of CD40 on different lysates with Mouse anti-CD40 antibody (EM1901-40) at 1/1,000 dilution.

Lane 1: Raji cell lysate
Lane 2: HeLa cell lysate (negative)
Lane 3: Daudi cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 31 kDa
Observed band size: 40 kDa

Exposure time: Lane 1-2: 1 minute 40 seconds; Lane 3: 3 minutes; ECL: K1801;

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 (EM1901-40) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
EM1901-40_2.jpg Fig2: ICC staining of CD40 in MG-63 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 (EM1901-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
EM1901-40_3.jpg Fig3: ICC staining of CD40 in SiHa 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 (EM1901-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
EM1901-40_4.jpg Fig4: ICC staining of CD40 in SKOV-3 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 (EM1901-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
EM1901-40_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-CD40 antibody (EM1901-40) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-40) at 1/1,000 dilution for 1 hour 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.