MA5-24164

antibody from Invitrogen Antibodies

Targeting: CEACAM6 CD66c, NCA

Western blot (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-24164

Provider

Invitrogen Antibodies

Product name

CEACAM6 Monoclonal Antibody (439424)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs and Western blots, no cross-reactivity with recombinant human CEACAM-1, -3, or -5 is observed.

Antibody clone number

439424

Concentration

0.5 mg/mL

References

Submitted references

Biological Functions and Identification of Novel Biomarker Expressed on the Surface of Breast Cancer-Derived Cancer Stem Cells via Proteomic Analysis.

Koh EY, You JE, Jung SH, Kim PH
Molecules and cells 2020 Apr 30;43(4):384-396

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CEACAM6 Monoclonal Antibody (439424) (Product # MA5-24164) and a 80 kDa band corresponding to Carcinoembryonic antigen-related cell adhesion molecule 6 was observed across tissues tested. Tissue extracts (50 µg lysate) of Mouse Liver (Lane 1), Mouse Lung (Lane 2), Mouse Ovary (Lane 3) and Mouse Heart (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of HEK293 human embryonic kidney cell line transfected with (A) human CEACAM-6/CD66c or (B) irrelevant transfectants and eGFP was stained with Mouse Anti-Human CEACAM-6/CD66c Monoclonal Antibody (Product # MA5-24164) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody. Quadrant markers were set based on control antibody staining.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of CEACAM6 in HEK293 human embryonic kidney cell line transfected with (A) human CEACAM-6/CD66c or (B) irrelevant transfectants and eGFP. Samples were incubated in CEACAM6 monoclonal antibody (Product # MA5-24164) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody. Quadrant markers were set based on control antibody staining.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of CEACAM6 in Human whole blood granulocytes. Samples were incubated in CEACAM6 monoclonal antibody (Product # MA5-24164) or isotype control antibody followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab)2 Secondary Antibody.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 7 The expression of CD66c at the protein level in cultured BCSCs in a time-dependent manner. The expression of CD66c protein increased in a time-dependent manner in BSCSs compared to MCF-7 cells, in agreement with the transcriptomic results. These results indicate that the expression of CD66c is related to the maintenance period in CSCs, but not in cancer cells. (A) Confirmation of the expression of CD66c protein by western blotting. (B) Quantitative graph of the western blot. Data are expressed as the mean +- SEM. *** P < 0.01 versus MCF-7 group. (C) Resulting peaks of the CD66c protein levels as a factor of time expressed on the surface of BCSCs in the form of a histogram according to FACS data. (D) Quantitative graph of the shifted peak in the FACS data. Data are expressed as the mean +- SEM. * P < 0.05 and *** P < 0.01 for MCF-7 versus BCSC (7 days) or BCSC (14 days); **P < 0.02 for BCSC (14 days) versus BCSC (21 days). (E) The images of CD66c expressed on the surface of the BCSCs according to cell binding assay. Data are expressed as the mean +- SEM. ** P < 0.02 versus MCF-7 group. (F) Identification of the characteristics of CSCs in BCSCs expressing CD66c by immunocytochemistry.