United Kingdom (UK)
Recombinant monoclonal antibody to CEACAM1. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma B3-17.
UniProt Accession Number of Target Protein: P13688
Alternative Name(s) of Target: CD66a; Carcinoembryonic antigen-related cell adhesion molecule 1; Biliary glycoprotein 1; BGP-1; BGPI
Immunogen: This antibody was raised by immunising mice with recombinant human CEACAM1.
Specificity: This antibody is specific for human CEACAM1, and recognises an epitope in the extracellular domain.
Application Notes: In flow cytometric analysis, this antibody has been used to detect CEACAM1 surface expression by human epithelial and endothelial cell lines, and microvesicles derived from these cells (Muturi et al, 2013), normal human bronchial epithelial (NHBE) cells (Klaile et al, 2013), CEACAM1-knockout human intestinal epithelial cells (Klaile et al, 2017) and human B-cell subpopulations (Khairnar et al, 2015; Seifert et al, 2015). It has also been used to detect CEACAM1 expression in ELISA analysis of transfected A549 epithelial cells (Singer et al, 2014). This antibody has been used in immunofluorescence analysis of NHBE cells (Klaile et al, 2013), as well as in the immunoprecipitation of CEACAM1 from NHBE cell culture supernatants (Klaile et al, 2013). Additionally, this antibody has been used in IHC staining of paraffin-embedded human lung tissue sections (Klaile et al, 2013).
Antibody first published in:
Klaile et al. Carcinoembryonic antigen (CEA)-related cell adhesion molecules are co-expressed in the human lung and their expression can be modulated in bronchial epithelial cells by non-typable Haemophilus influenzae, Moraxella catarrhalis, TLR3, and type I and II interferons. Respir Res. 2013 Aug 14;14:85 PMID:23941132
Note on publication:
Describes the original use of this antibody, in FC, IF an IP analysis, to assess CEACAM1 expression by normal human bronchial epithelial (NHBE) cells.
Immunofluorescence staining of HepG2 cells using anti-CEACAM1 antibody (Ab01369) B3-17 Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells immobilized on Shi-fix™ cover-slips and stained with the chimeric rabbit IgG version of B3-17 (Ab01369-23.0) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01369-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (Ab00102-23.0) followed by Alexa Fluor® 488 secondary antibody.
Flow-cytometry using the anti-CEACAM1 antibody B3-17 (Ab01369). HepG2 cells were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the rabbit IgG1 version of B3-17 (Ab01369-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer
Western Blot using anti-CEACAM1 antibody B3-17 (Ab01369) HepG2 cell lysate (A), human colon (B) and human liver (C) tissue lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of B3-17 (Ab01369-23.0) at 0.1 µg/ml, 0.1 µg/ml and 0.03 µg/ml, respectively, before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Western Blot using anti-CEACAM1 antibody B3-17 (Ab01369). HepG2(A) and human liver(B) tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric mouse IgG version of B3-17 (Ab01369-1.1) at 0.1 µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of HepG2 cells using anti-CEACAM1 (Ab01369) B3-17 Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells stained with the chimeric mouse IgG version of B3-17 (Ab01369-1.1) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01215-1.1, DAPI, merged channels and an isotype control. The isotype control was stained with anti-unknown antibody (Ab00178-1.1) followed by Alexa Fluor® 488 secondary antibody.
Western Blot using anti-CEACAM1 antibody B3-17 (Ab01369). HepG2(A) and human liver(B) tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric mouse IgG version of B3-17 (Ab01369-1.1) at 0.1 µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Flow cytometry using the Anti-CEACAM1 antibody B3-17 (Ab01369). HepG2 cells were stained with anti-unknown specificity antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG1 version of B3-17 (Ab01369-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.