HA751331

CD40 Recombinant Rabbit Monoclonal Antibody [PSH10-01]

cat.: HA751331

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Cynomolgus monkey, Pig
Applications:	IHC-P, IHC-Fr, IF-Cell, FC, IF-Tissue
Clonality:	Monoclonal
Clone number:	PSH10-01

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 31 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human CD40 aa 228-277 / 277.
Positive control:	Human spleen tissue, mouse spleen tissue, rat spleen tissue, Raji, A20, PC-12.
Subcellular location:	Cell membrane; Secreted.
Recommended Dilutions: IHC-P IHC-Fr IF-Cell FC IF-Tissue	1:1,000 1:1,000 1:50-1:500 1:1,000 1:500
Uniprot #:	SwissProt: P25942 Human \| P27512 Mouse Entrez Gene: 171369 Rat
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

	Fig1: Application: IHC-Fr Species: Mouse Site: Spleen Sample: Frozen section Antibody concentration: 1:500 Antigen retrieval: Not required
	Fig2: Application: IHC-Fr Species: Rat Site: Spleen Sample: Frozen section Antibody concentration: 1:500 Antigen retrieval: Not required
	Fig3: Application: IF-Tissue Species: Mouse Site: spleen Sample: Paraffin-embedded section Antibody concentration: 1/500
	Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD40 antibody (HA751331) 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 (HA751331) 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.

	Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-CD40 antibody (HA751331) 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 (HA751331) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-CD40 antibody (HA751331) 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 (HA751331) 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.
	Fig7: Immunocytochemistry analysis of Raji cells labeling CD40 with Rabbit anti-CD40 antibody (HA751331) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD40 antibody (HA751331) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Fig8: Immunocytochemistry analysis of A20 cells labeling CD40 with Rabbit anti-CD40 antibody (HA751331) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD40 antibody (HA751331) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Fig9: Immunocytochemistry analysis of PC-12 cells labeling CD40 with Rabbit anti-CD40 antibody (HA751331) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD40 antibody (HA751331) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Fig10: Flow cytometric analysis of A20 cells labeling CD40.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751331, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. 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”.