Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [2]
- Flow cytometry [1]
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Validation data
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- Product number
- MA1-17761 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD66 (CEACAM) Monoclonal Antibody (D14HD11)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- This antibody recognizes native human CEACAM1, 3, 4, 5, 6 protein expressed in transfected mammalian cells.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- D14HD11
- Vial size
- 100 µg
- Concentration
- 2 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-CEACAM Pan Monoclonal Antibody (D14HD11) (Product # MA1-17761) and a 150 kDa band corresponding to Carcinoembryonic antigen-related cell adhesion molecule 5 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), HCT 116 (Lane 2), MDA-MB-231 (Lane 3), HT-29 (Lane 4), HT-29 treated with IL-6 (100 ng/mL, 48 Hrs) (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 Dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry microscopy of CD66 (CEACAM) in CHO cells using CD66 (CEACAM) Monoclonal Antibody (Product # MA1-17761). CHO cells were transfected with an expression vector encoding CEACAM1 (A). Untransfected CHO parental cells served as negative control (B). Binding was visualized with a FITC-conjugated secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Carcinoembryonic antigen-related cell adhesion molecule 5 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CEACAM Pan Monoclonal Antibody (D14HD11) (Product # MA1-17761) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Cytoplasm localization. Panel e represents MDA-MB-231. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of CEACAM 1,3,4,5,6 in BOSC23 cells using a CD66 (CEACAM) Monoclonal Antibody (Product # MA1-17760) at a 1.2 µg/10^6 cells dilution, detected with a PE-conjugated secondary antibody, as shown in red. Isotype control antibody is shown in black.