Recombinant Anti-CEACAM1 antibody [EPR26490-88] (ab300061)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26490-88] to CEACAM1
- Suitable for: Flow Cyt, IHC-P, WB, IP, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CEACAM1 antibody [EPR26490-88]
See all CEACAM1 primary antibodies -
Description
Rabbit monoclonal [EPR26490-88] to CEACAM1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt, IHC-P, WB, IP, ICC/IFmore details
Unsuitable for: IHC-Fr -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Recombinant fragment corresponding to Human CEACAM1 aa 1-450.
Database link: P13688 -
Positive control
- WB: Wild-type A549, HepG2, Human kidney, Human colon, A-204, SW480, His-tagged human CEACAM-1 recombinant protein aa35-428 recombinant protein lysates. IHC-P: Human: colon, endometrium, liver and hepatic carcinoma tissues ICC/IF: A549 cells. Flow cyt.: Wild-type A549 cell. IP: A549 cell.
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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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR26490-88 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab300061 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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Flow Cyt |
1/500.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 57 kDa.
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IP |
1/30.
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ICC/IF |
1/500.
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Notes |
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Flow Cyt
1/500. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 57 kDa. |
IP
1/30. |
ICC/IF
1/500. |
Target
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Sequence similarities
Belongs to the immunoglobulin superfamily. CEA family.
Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Cellular localization
Secreted; Cell membrane and Cell membrane. Localizes to sites of cell-cell contact. - Information by UniProt
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Database links
- Entrez Gene: 634 Human
- Omim: 109770 Human
- SwissProt: P13688 Human
- Unigene: 512682 Human
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Form
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. -
Alternative names
- Antigen CD66 antibody
- BGP 1 antibody
- BGP antibody
see all
Images
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All lanes : Anti-CEACAM1 antibody [EPR26490-88] (ab300061) at 1/1000 dilution
Lane 1 : Wild-type A549 (ab255450) whole cell lysate
Lane 2 : CEACAM1 knockout A549 (ab267047) whole cell lysate
Lane 3 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) at 1/10000 dilution
Predicted band size: 57 kDa
Observed band size: 100-180 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab300061 observed at 100-180 kDa. Red - loading control ab8245 observed at 36 kDa.
ab300061 Anti-CEACAM1 antibody [EPR26490-88] was shown to specifically react with CEACAM1 in wild-type A549 cells (ab255450). Loss of signal was observed when knockout cell line (ab267047) was used. Wild-type and CEACAM1 knockout samples were subjected to SDS-PAGE. ab300061 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging. -
All lanes : Anti-CEACAM1 antibody [EPR26490-88] (ab300061) at 1/1000 dilution
Lane 1 : Human kidney tissue lysate
Lane 2 : Human colon tissue lysate
Lane 3 : A-204 (human muscle rhabdomyosarcoma) whole cell lysate
Lane 4 : SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L)
Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 100-180 kDa why is the actual band size different from the predicted?Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: Lanes 1-3:10 seconds; Lane 4: 48 seconds.
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All lanes : Anti-CEACAM1 antibody [EPR26490-88] (ab300061) at 1/1000 dilution
Lane 1 : His-tagged human CEACAM-1 recombinant protein aa (aa35-428) 20ng
Lane 2 : His-tagged human CEACAM-5/CD66e recombinant protein (aa35-685) 20ng
Lane 3 : His-tagged human CEACAM-6/CD66c recombinant protein (aa35-320) 20ng
Lane 4 : His-tagged human CEACAM-8/CD66b recombinant protein(aa35-320) 20ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 100-180 kDa why is the actual band size different from the predicted?
Exposure time: 48 secondsBlocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: Lanes 1-3:10 seconds; Lane 4: 48 seconds.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labelling CEACAM1 with ab300061 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human colon (PMID: 8543780). The section was incubated with ab300061 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody (ab300061) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded human endometrium tissue labelling CEACAM1 with ab300061 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human endometrium (PMID: 9626043). The section was incubated with ab300061 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody (ab300061) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded human liver tissue labelling CEACAM1 with ab300061 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human liver (PMID: 27377843). The section was incubated with ab300061 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody (ab300061) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded human hepatic carcinoma tissue labelling CEACAM1 with ab300061 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human hepatic carcinoma (PMID: 27377843). The section was incubated with ab300061 for 30 mins at room temperature The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody (ab300061) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized CEACAM1 KO A549 (CEACAM1 knockout human lung carcinoma epithelial cell) (ab267047) cells lebelling CEACAM1 with ab300061 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membrane and cytoplasmic staining in parental A549 cells, no staining in CEACAM1 KO A549 cells.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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Flow cytometric analysis of Wild-type A549 (Human lung carcinoma epithelial cell, Right)/ CEACAM1 knockout A549 cells (Left) cells labelling CEACAM1 with ab300061 at 1/500 dilution/ Left and Right compared with a isotype control. Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Positive staining on A549 cells (ab255450), while no staining on CEACAM1 knockout A549 cells (ab267047).
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CEACAM1 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) whole cell lysate 10 µg with ab300061 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab300061 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: A549 (human lung carcinoma epithelial cell) whole cell lysate 10 µg (Input)
Lane 2: ab300061 IP in A549 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300061 in A549 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
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 (0)
ab300061 has not yet been referenced specifically in any publications.