Recombinant Anti-CEACAM5 + CD66b + CEACAM1 + CEACAM6 antibody [EPR20721] (ab207718)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20721] to CEACAM5 + CD66b + CEACAM1 + CEACAM6
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CEACAM5 + CD66b + CEACAM1 + CEACAM6 antibody [EPR20721] -
Description
Rabbit monoclonal [EPR20721] to CEACAM5 + CD66b + CEACAM1 + CEACAM6 -
Host species
Rabbit -
Specificity
ab207718 cross-reacts with CEACAM 1, 6 and 8. There are other CEACAM family members but we only examined cross-reactivity with those having >55% homology with CEACAM5 following a BLAST search with the immunogen. -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: His-GST-tagged human CEACAM5 recombinant protein (aa238-686), His-tagged human CEACAM6 recombinant protein (aa35-326), CEACAM8 recombinant protein (aa35-335) and CEACAM1 recombinant protein (aa35-428). MCF7, LoVo and Caco-2 whole cell lysate. Human colon lysate. IHC-P: Human colon and colon cancer tissue. Human spleen tissue, HEK-293T cells transfected with a CEACAM3, CEACAM5 and CEACAM8 expression vector containing a his tag. ICC/IF, Flow Cyt: MCF7 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20721 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry reagents
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab207718 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Predicted molecular weight: 76 kDa.
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IHC-P |
1/5000 - 1/40000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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Flow Cyt |
1/500.
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Notes |
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WB
1/1000. Predicted molecular weight: 76 kDa. |
IHC-P
1/5000 - 1/40000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
Flow Cyt
1/500. |
Target
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Cellular localization
CEACAM5: Cell membrane. CD66b: Cell membrane. CEACAM1: Secreted; Cell membrane and Cell membrane. Localizes to sites of cell-cell contact. CEACAM6: Cell membrane. -
Database links
- Entrez Gene: 1048 Human
- Entrez Gene: 1088 Human
- Entrez Gene: 4680 Human
- Entrez Gene: 634 Human
- Omim: 109770 Human
- Omim: 114890 Human
- Omim: 163980 Human
- Omim: 615747 Human
see all -
Form
CEACAM1: There are 11 isoforms produced by alternative splicing. Isoform 1 = BGPa; CEACAM1-4L; TM1-CEA; Isoform 2 = BGPg; CEACAM1-4C1; Isoform 3 = BGPh; CEACAM1-3; Isoform 4 = BGPi; CEACAM1-3C2; Isoform 5 = BGPy; CEACAM1-3AL; Isoform 6 = BGPb; CEACAM1-3L; TM2-CEA; Isoform 7 = BGPx; CEACAM1-1L; Isoform 8 = BGPc; CEACAM1-4S; TM3-CEA; Isoform 9 = BGPz; CEACAM1-3AS and Isoform 11 = BGPd; CEACAM1-3S.
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carcino Embryonic Antigen CEA antibody [EPR20721] (ab207718)
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Carcino Embryonic Antigen CEA with ab207718 at 1/40,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human colon (PMID: 25042385) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
All lanes : Anti-CEACAM5 + CD66b + CEACAM1 + CEACAM6 antibody [EPR20721] (ab207718) at 1/1000 dilution
Lane 1 : His-GST-tagged human CEACAM5 recombinant protein (aa238-686) 10 ng
Lane 2 : His-tagged human CEACAM6 recombinant protein (aa35-326) 10 ng
Lane 3 : His-tagged human CEACAM8 recombinant protein (aa35-335) 10 ng
Lane 4 : His-tagged human CEACAM1 recombinant protein (aa35-428) 10 ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 76 kDa
Observed band size: 30,50,75 kDa why is the actual band size different from the predicted?
Additional bands at: 32 kDa (possible post-translational modification)Blocking/ Dilution buffer and concentration: 5% NFDM/TBST.
Exposure times:
Lanes 1-3: 10 seconds
Lane 4: 4 minutes
Observed MW: 50, 30, 32, 75 kDa.
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
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All lanes : Anti-CEACAM5 + CD66b + CEACAM1 + CEACAM6 antibody [EPR20721] (ab207718) at 1/1000 dilution
Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 76 kDa
Observed band size: 110-150,75-90 kDa why is the actual band size different from the predicted?
Exposure time: 58 secondsBlocking/ Dilution buffer and concentration: 5% NFDM/TBST.
Observed MW: 110-150, 75-90 kDa.
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
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All lanes : Anti-CEACAM5 + CD66b + CEACAM1 + CEACAM6 antibody [EPR20721] (ab207718) at 1/1000 dilution
Lane 1 : Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Lane 2 : SW480 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Lane 3 : Human colon lysate at 20 µg
Lane 4 : LoVo (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 76 kDa
Observed band size: 110-150,75-90 kDa why is the actual band size different from the predicted?Blocking/ Dilution buffer and concentration: 5% NFDM/TBST.
Exposure times:
Lanes 1-2: 3 minutes
Lanes 3-4: 10 seconds
Observed MW 110-150, 75-90 kDa.
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument. The expression profile observed is consistent with differential glycosylation of this protein (PMID: 1717152).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carcino Embryonic Antigen CEA antibody [EPR20721] (ab207718)
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Carcino Embryonic Antigen CEA with ab207718 at 1/40,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human colon cancer (PMID: 25042385) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carcino Embryonic Antigen CEA antibody [EPR20721] (ab207718)
Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Carcino Embryonic Antigen CEA with ab207718 at 1/40,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining of spleen-infiltrating neutrophils (PMID: 22064717 and 19077207) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunocytochemistry/ Immunofluorescence - Anti-Carcino Embryonic Antigen CEA antibody [EPR20721] (ab207718)
Immunofluorescent analysis of 100% methanol fixed MCF7 (human breast adenocarcinoma epithelial cell) cells labeling Carcino Embryonic Antigen CEA with ab207718 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing membranous staining on MCF7 cell line.
Negative control for cells: PANC-1 .
Counterstained with ab195889 anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red) and DAPI (blue).
The negative control is the secondary antibody only.
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Flow cytometric analysis of PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) (Left) / MCF7 (human breast adenocarcinoma epithelial cell) (Right) labeling Carcino Embryonic Antigen CEA with ab207718 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Cells were not fixed or permeabilized. Gated on total viable cells.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 + CD66b + CEACAM1 + CEACAM6 antibody [EPR20721] (ab207718)
Immunohistochemistry analysis of paraffin-embedded (A) HEK-293T cells transfected with a CEACAM1 expression vector containing a his tag, (B) HEK-293T cells transfected with a CEACAM3 expression vector containing a his tag, (C) HEK-293T cells transfected with a CEACAM5 expression vector containing a his tag, (D) HEK-293T cells transfected with a CEACAM6 expression vector containing a his tag, (E) HEK-293T cells transfected with a CEACAM8 expression vector containing a his tag, (F) HEK-293T cells transfected with empty vector containing a his tag, sections labelling CEACAM5 + CD66b + CEACAM1 + CEACAM6 with ab207718 at 1/5000 dilution. The section was incubated with ab207718 for 30 mins at room temperature. LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Positive stainng on (B) HEK-293T cells transfected with a CEACAM3 expression vector containing a his tag, (C) HEK-293T cells transfected with a CEACAM5 expression vector containing a his tag, (E) HEK-293T cells transfected with a CEACAM8 expression vector containing a his tag. Nearly no staining on (A) HEK-293T cells transfected with a CEACAM1 expression vector containing a his tag, (D) HEK-293T cells transfected with a CEACAM6 expression vector containing a his tag and (F) HEK-293T cells transfected with empty vector containing a his tag. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab207718 has been referenced in 1 publication.
- Liu X et al. RNA-binding protein with serine-rich domain 1 regulates microsatellite instability of uterine corpus endometrial adenocarcinoma. Clinics (Sao Paulo) 76:e3318 (2021). PubMed: 34817046