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CD146 Human Recombinant Antibody

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  • CD146 Human Recombinant Antibody
  • Capture ELISA of CD146/MCAM: As validation of the antibody, 1ug/ml of CD146/MCAM antigen or control protein (BSA and Fc) were adsorbed onto maxisorp plates, blocked with PBS + 0.5% BSA (PB), and washed 3 times with PBS + 0.05% Tween-20 (PT); 0.2µg /ml anti-CD146/MCAM hIgG1 antibody in PBS for 30 min, shaking RT, washed 6 times with PT.  Anti-human kappa-HRP (1:5000 in PB) was incubated for 30 min, shaking RT, washed 6 times with PT. Colorimetric HRP reagents allow for absorbance readings at 450 nm. Capture ELISA of CD146/MCAM: As validation of the antibody, 1ug/ml of CD146/MCAM antigen or control protein (BSA and Fc) were adsorbed onto maxisorp plates, blocked with PBS + 0.5% BSA (PB), and washed 3 times with PBS + 0.05% Tween-20 (PT); 0.2µg /ml anti-CD146/MCAM hIgG1 antibody in PBS for 30 min, shaking RT, washed 6 times with PT. Anti-human kappa-HRP (1:5000 in PB) was incubated for 30 min, shaking RT, washed 6 times with PT. Colorimetric HRP reagents allow for absorbance readings at 450 nm.
  • Flow Cytometry of CD146/MCAM: Hela cells were collected using mild EDTA solution and washed in PBS. 5x105 cells were incubated with or without 1 ug of anti-CD146/MCAM Fab antibody in 100 ulL PBS + 0.1% BSA (PB) for 30 min on ice, then washed twice with cold PB. Samples were then incubated with Goat anti-Human Fab-488 (Jackson ImmunoResearch, diluted 1:1000 PB) for 15-30 min, washed twice with cold PBS and fixed with 2% paraformaldehyde and filtered prior to reading on a FACs machine. Flow Cytometry of CD146/MCAM: Hela cells were collected using mild EDTA solution and washed in PBS. 5x105 cells were incubated with or without 1 ug of anti-CD146/MCAM Fab antibody in 100 ulL PBS + 0.1% BSA (PB) for 30 min on ice, then washed twice with cold PB. Samples were then incubated with Goat anti-Human Fab-488 (Jackson ImmunoResearch, diluted 1:1000 PB) for 15-30 min, washed twice with cold PBS and fixed with 2% paraformaldehyde and filtered prior to reading on a FACs machine.
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CD146 Human Recombinant Antibody

SKU:
MM-0302

USD 350.00 /100 µg

Clonality: 
Monoclonal
Host: Human; made using CCAB phage display library
Reactivity: Human
Application: Flow Cytometry

QTY:

OVERVIEW

Target: 
Human CD146
Target background: 
This protein plays a role in cell adhesion, and in cohesion of the endothelial monolayer at intercellular junctions in vascular tissue. Its expression may allow melanoma cells to interact with cellular elements of the vascular system, enhancing hematogeneous tumour spread. [Provided by UniProt]
Target alias: 
MCAM, MUC18
Immunogen: 
ECD of recombinant CD146, aa: Met1-Gly559 
Specificity: This antibody recognizes the ECD of CD146 (aa: Met1-Gly559 )
Clone ID: 
CR - 003 (generated by CCAB)
Isotype: 
hIgG1; also available as mIgG1, scIgG1, schmIgG2a
Preservative: 
None
Format: 
Lyophilized protein A purified in PBS pH7.4
Recommend starting dilution: 
Reconstitute with deionized water. Optimal dilution has to be determined by the user.
Limitations: 
Research Use Only; Manufactured by CCAB under a license using Life Technologies proprietary cell lines.
Storage: 
Lyophilized antibodies can be kept at 4ºC for up to 3 months and should be kept at -20ºC for long-term storage (2 years). To avoid freeze-thaw cycles, reconstituted antibodies should be aliquoted before freezing for long-term (1 year) storage (-80ºC) or kept at 4ºC for short-term usage (2 months). For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made with the assay buffer. After the maximum long-term storage period (2 years lyophilized or 1 year reconstituted) antibodies should be tested in your assay with a standard sample to verify if you have noticed any decrease in their efficacy.