Anti-Carcino Embryonic Antigen CEA 抗体 [EPCEAR7] (ab133633)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPCEAR7] to CEACAM5
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7]
CEACAM5 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPCEAR7] to CEACAM5 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-P, ICC/IFmore details -
種交差性
交差種: Human -
免疫原
Full length native protein (purified) corresponding to Human Carcino Embryonic Antigen CEA.
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ポジティブ・コントロール
- WB: Human colon cancer lysate, His-tagged human CEACAM5 recombinant protein (aa 35-586), BxPC-3 and MCF7 whole cell lysates unboiled. IHC-P: Human colon carcinoma tissue, human gastric carcinoma tissue, HEK-293T cells transfected with a CEACAM5 expression vector containing a his tag and HEK-293T cells transfected with a CEACAM6 expression vector containing a his tag. ICC/IF: MCF7 cells.
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特記事項
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
製品の特性
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製品の状態
Liquid -
保存方法
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. Stable for 12 months at -20°C. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPCEAR7 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Isotype control
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab133633の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
1/1000 - 1/10000. Detects a band of approximately 180-200 kDa (predicted molecular weight: 76 kDa).
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IHC-P |
1/2000 - 1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. ab133633 is also cross-reactIve with human CEACAM6 in IHC. |
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ICC/IF |
1/50 - 1/500.
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特記事項 |
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WB
1/1000 - 1/10000. Detects a band of approximately 180-200 kDa (predicted molecular weight: 76 kDa). |
IHC-P
1/2000 - 1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. ab133633 is also cross-reactIve with human CEACAM6 in IHC. |
ICC/IF
1/50 - 1/500. |
ターゲット情報
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機能
Cell surface glycoprotein that plays a role in cell adhesion and in intracellular signaling. Receptor for E.coli Dr adhesins. -
組織特異性
Found in adenocarcinomas of endodermally derived digestive system epithelium and fetal colon. -
配列類似性
Belongs to the immunoglobulin superfamily. CEA family.
Contains 7 Ig-like (immunoglobulin-like) domains. -
翻訳後修飾
Complex immunoreactive glycoprotein with a MW of 180 kDa comprising 60% carbohydrate. -
細胞内局在
Cell membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 1048 Human
- Omim: 114890 Human
- SwissProt: P06731 Human
- Unigene: 709196 Human
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別名
- Carcinoembryonic antigen antibody
- Carcinoembryonic antigen-related cell adhesion molecule 5 antibody
- CD66e antibody
see all
画像
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All lanes : Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : CEACAM5 knockout A549 cell lysate
Lane 3 : MCF7 (unboiled) cell lysate
Lane 4 : PANC-1 (unboiled) cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 76 kDa
Observed band size: 160-180 kDa why is the actual band size different from the predicted?Western blot: Anti-CEACAM5 antibody [EPCEAR7] (ab133633) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab133633 was shown to bind specifically to CEACAM5. A band was observed at 160-180 kDa in wild-type A549 cell lysates with no signal observed at this size in CEACAM5 knockout cell line. To generate this image, wild-type and CEACAM5 knockout A549 unboiled cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633) at 1/1000 dilution
Lane 1 : BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate boiled
Lane 2 : BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate unboiled
Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate boiled
Lane 4 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate unboiled
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 76 kDa
Observed band size: 180-200 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as a loading control.
We recommend not to boil the sample after lysis to get desirable WB result.
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Confocal image showing membranous staining in MCF7 cell line.
Immunocytochemistry/Immunofluorescence analysis of MCF7 (human breast adenocarcinoma cell line) cells labelling carcino embryonic antigen (CEA) with ab133633 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/10000 dilution was used as the secondary antibody. The cells were counterstained with ab195889, a anti-alpha tubulin antibody at 1/200 dilution. DAPI was used as a nuclear counterstain.
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling Carcino Embryonic Antigen CEA with Purified ab133633 at 1:3000 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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All lanes : Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633) at 1/1000 dilution
Lane 1 : His-tagged human CEACAM1 recombinant protein (aa 35-428)
Lane 2 : His-tagged human CEACAM3 recombinant protein (aa 35-252)
Lane 3 : His-tagged human CEACAM5 recombinant protein (aa 35-586)
Lane 4 : His-tagged human CEACAM6 recombinant protein (aa 35-320)
Lane 5 : His-tagged human CEACAM8 recombinant protein (aa 35-320)
Lysates/proteins at 0.02 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: 180 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBSTThis antibody does not cross-react with human CEACAM1, CEACAM3, CEACAM6, and CEACAM8.
This blot was developed using a high sensitivity ECL substrate.
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Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633) at 1/1000 dilution + Human colon cancer lysate at 10 µg
Secondary
Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Predicted band size: 76 kDa
Observed band size: 180-200 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human gastric carcinoma tissue sections labeling Carcino Embryonic Antigen CEA with Purified ab133633 at 1:3000 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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ab133633 showing positive staining in Normal colon tissue.
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Immunohistochemical analysis of paraffin embedded Human colon carcinoma tissue labelling Carcino Embryonic Antigen CEA with ab133633 at 1/250.
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Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633) at 0.4 µg/ml (Purified) + Human colon cancer lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 76 kDaBlocking and diluting buffer: 5% NFDM/TBST
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ab133633 showing positive staining in Stomach adenocarcinoma tissue.
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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 and (F) HEK-293T cells transfected with empty vector containing a his tag, sections labelling Carcino Embryonic Antigen CEA with ab133633 at 1/2000 dilution. LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Sections were counterstained with Hematoxylin. The section was incubated with ab133633 for 30 mins at room temperature. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Positive staining on HEK-293T cells transfected with a CEACAM5 expression vector containing a his tag (Image C), and HEK-293T cells transfected with a CEACAM6 expression vector containing a his tag (Image D). No staining on other cell pellet tested. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.IHC Note: ab133633 is also cross-reactIve with human CEACAM6 in IHC.
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ab133633 showing negative staining in Skeletal muscle tissue.
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ab133633 showing negative staining in Normal tonsil tissue.
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ab133633 showing negative staining in Thyroid gland carcinoma tissue.
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ab133633 showing negative staining in Hepatocellular carcinoma tissue.
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ab133633 showing negative staining in Glioma tissue.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (8)
ab133633 は 8 報の論文で使用されています。
- Tan D et al. Screening of an individualized treatment strategy for an advanced gallbladder cancer using patient-derived tumor xenograft and organoid models. Front Oncol 12:1043479 (2022). PubMed: 36591461
- Miyamoto Y et al. Epidermal growth factor receptor-targeted molecular imaging of colorectal tumors: Detection and treatment evaluation of tumors in animal models. Cancer Sci 110:1921-1930 (2019). PubMed: 30973663
- Cheng M et al. Comprehensive analysis of marker gene detection and computed tomography for the diagnosis of human lung cancer. Oncol Lett 16:4400-4406 (2018). PubMed: 30250539
- Ramos-Gomes F et al. Single- and two-photon imaging of human micrometastases and disseminated tumour cells with conjugates of nanobodies and quantum dots. Sci Rep 8:4595 (2018). WB . PubMed: 29545609
- Gao J et al. Contrast-enhanced magnetic resonance imaging with a novel nano-size contrast agent for the clinical diagnosis of patients with lung cancer. Exp Ther Med 15:5415-5421 (2018). PubMed: 29904421
- Tian HW et al. Multiple pulmonary metastases with halo-sign from malignant mixed Müllerian tumors. Oncol Lett 14:6645-6649 (2017). PubMed: 29151911
- Lugini L et al. Exosomes from human colorectal cancer induce a tumor-like behavior in colonic mesenchymal stromal cells. Oncotarget 7:50086-50098 (2016). PubMed: 27418137
- Xin H et al. Establishment and characterization of 7 novel hepatocellular carcinoma cell lines from patient-derived tumor xenografts. PLoS One 9:e85308 (2014). IHC-P ; Human . PubMed: 24416385