Anti-CEACAM1 antibody [EPR4049] (ab108397)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4049] to CEACAM1
- Suitable for: WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-CEACAM1 antibody [EPR4049]
See all CEACAM1 primary antibodies -
Description
Rabbit monoclonal [EPR4049] to CEACAM1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanIP HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal liver, A549, SW480, and HepG2 cell lysates; IHC-P: Human kidney clear cell carcinoma tissue. IP: SW480 cells
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
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
EPR4049 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-CEACAM1 antibody [EPR4049] (ab108397) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : CEACAM1 knockout A549 cell lysate
Lane 3 : A549 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 58 kDa
Observed band size: 100-180 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab108397 observed at 100-180 kDa. Red - loading control ab8245 observed at 36 kDa.
ab108397 Anti-CEACAM1 antibody [EPR4049] was shown to specifically react with CEACAM1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267046 (knockout cell lysate ab257388) was used. Wild-type and CEACAM1 knockout samples were subjected to SDS-PAGE. ab108397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 antibody [EPR4049] (ab108397)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human clear cell carcinoma of kidney tissue sections labeling CEACAM1 with Purified ab108397 at 1:80 dilution (3.3 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-CEACAM1 antibody [EPR4049] (ab108397) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : CEACAM1 knockout A549 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 58 kDa
Observed band size: 100-180 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab108397 observed at 100-180 kDa. Red - loading control ab8245 observed at 36 kDa.
ab108397 Anti-CEACAM1 antibody [EPR4049] was shown to specifically react with CEACAM1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267047 (knockout cell lysate ab257389) was used. Wild-type and CEACAM1 knockout samples were subjected to SDS-PAGE. ab108397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-CEACAM1 antibody [EPR4049] (ab108397) at 1/10000 dilution (Purified)
Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
Lane 2 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
Lane 3 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 58 kDa
Observed band size: 100-180 kDa why is the actual band size different from the predicted?The smear and multiple bands are consistent with what have been described in the literature PMID: 17071610, 26448580 and 30050594, due to a variety of splice variants and/or differential glycosylation
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ab108397 (purified) at 1:20 dilution (1.5µg) immunoprecipitating CEACAM1 in SW480 whole cell lysate.
Lane 1 (input): SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab108397 & SW480 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108397 in SW480 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 antibody [EPR4049] (ab108397)
Unpurified ab108397 at 1/50 dilution staining CEACAM1 in Human kidney tissue by Immunohistochemistry, Paraffin-embedded tissue.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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