Product Info Summary
SKU: | M01683-3 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human |
Host: | Mouse |
Application: | Flow Cytometry, IHC, IHC-F, WB |
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Product info
Product Name
Anti-CD146/MCAM Antibody Picoband™ (monoclonal, 4C12)
View all MCAM/CD146 Antibodies
SKU/Catalog Number
M01683-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-CD146/MCAM Antibody Picoband™ (monoclonal, 4C12) catalog # M01683-3. Tested in Flow Cytometry, IHC, IHC-F, WB applications. This antibody reacts with Human.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-CD146/MCAM Antibody Picoband™ (monoclonal, 4C12) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01683-3)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
4C12
Isotype
Mouse IgG2a
Immunogen
E.coli-derived human CD146 recombinant protein (Position: H59-A401). Human CD146 shares 73% amino acid (aa) sequence identity with both mouse and rat CD146.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M01683-3 is reactive to MCAM in Human
Applications
M01683-3 is guaranteed for Flow Cytometry, IHC, IHC-F, WB Boster Guarantee
Observed Molecular Weight
120 kDa
Calculated molecular weight
71.607kDa
Background of MCAM/CD146
CD146 (cluster of differentiation 146), also known as the melanoma cell adhesion molecule (MCAM) or cell surface glycoprotein MUC18, is a 113kDa cell adhesion molecule currently used as a marker for endothelial cell lineage. MCAM, a member of the immunoglobulin superfamily, is homologous to several cell adhesion molecules and is associated with tumor progression and the development of metastasis in human malignant melanoma. By radiation hybrid analysis, this gene is mapped to chromosome 11q23.3. MCAM has been demonstrated to appear on a small subset of T and B lymphocytes in the peripheral blood of healthy individuals. MCAM has been seen as a marker for mesenchymal stem cells isolated from multiple adult and fetal organs, and its expression may be linked to multipotency mesenchymal stem cells with greater differentiation potential express higher levels of MCAM on the cell surface.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of CD146/MCAM using anti-CD146/MCAM antibody (M01683-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human A375 whole cell lysates;
Lane 2: human Hela whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-CD146/MCAM antigen affinity purified monoclonal antibody (Catalog # M01683-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for CD146/MCAM at approximately 120KD. The expected band size for CD146/MCAM is at 72KD.
Click image to see more details
Figure 2. IHC analysis of CD146/MCAM using anti-CD146/MCAM antibody (M01683-3).
CD146/MCAM was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CD146/MCAM Antibody (M01683-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of CD146/MCAM using anti-CD146/MCAM antibody (M01683-3).
CD146/MCAM was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CD146/MCAM Antibody (M01683-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of CD146/MCAM using anti-CD146/MCAM antibody (M01683-3).
CD146/MCAM was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CD146/MCAM Antibody (M01683-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of CD146/MCAM using anti-CD146/MCAM antibody (M01683-3).
CD146/MCAM was detected in frozen section of human placenta tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CD146/MCAM Antibody (M01683-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 6. Flow Cytometry analysis of SiHa cells using anti-CD146/MCAM antibody (M01683-3).
Overlay histogram showing SiHa cells stained with M01683-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-CD146/MCAM Antibody (M01683-3, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For MCAM (Source: Uniprot.org, NCBI)
Gene Name
MCAM
Full Name
Cell surface glycoprotein MUC18
Weight
71.607kDa
Alternative Names
CD146 antigen; CD146; cell surface glycoprotein MUC18; Cell surface glycoprotein P1H12; L-Gicerin; MCAM; melanoma adhesion molecule; melanoma cell adhesion moleculeS-endo 1 endothelial-associated antigen; Melanoma-associated antigen A32; Melanoma-associated antigen MUC18; MUC18; MUC18Gicerin MCAM CD146, HEMCAM, METCAM, MUC18, MelCAM melanoma cell adhesion molecule cell surface glycoprotein MUC18|Gicerin|S-endo 1 endothelial-associated antigen|cell surface glycoprotein P1H12|melanoma adhesion molecule|melanoma-associated antigen A32|melanoma-associated antigen MUC18
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on MCAM, check out the MCAM Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for MCAM: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-CD146/MCAM Antibody Picoband™ (monoclonal, 4C12) (M01683-3)
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