ET7107-91

CD146 Recombinant Rabbit Monoclonal Antibody [JB42-35]

cat.: ET7107-91

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, FC, IF-Cell, IF-Tissue
Clonality:	Monoclonal
Clone number:	JB42-35

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: 72 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within C-terminal human CD146 (Cytoplasmic).
Positive control:	SiHa cell lysates, Hela, HUVEC, rat brain tissue, human tonsil tissue, mouse brain tissue.
Subcellular location:	Membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:500-1:1,000 1:100-1:500 1:100-1:500 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P43121 Human \| Q8R2Y2 Mouse \| Q9EPF2 Rat
Alternative names:	A32 antigen CD 146 CD146 CD146 antigen Cell surface glycoprotein MUC18 Cell surface glycoprotein P1H12 Gicerin Mcam Melanoma adhesion molecule Melanoma associated antigen A32 Melanoma associated antigen MUC18 Melanoma associated glycoprotein MUC18 Melanoma cell adhesion molecule Melanoma-associated antigen A32 Melanoma-associated antigen MUC18 MelCAM MUC 18 MUC18 MUC18_HUMAN S endo 1 S endo 1 endothelial associated antigen S-endo 1 endothelial-associated antigen

Images

	Fig1: Western blot analysis of CD146 on SiHa cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7107-91, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
	Fig2: ICC staining of CD146 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-91, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig3: ICC staining of CD146 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-91, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-CD146 antibody. 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 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7107-91, 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.

	Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD146 antibody. 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 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7107-91, 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-CD146 antibody. 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 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7107-91, 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.
	Fig7: Flow cytometric analysis of CD146 was done on HUVEC cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-91, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-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”.