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使用敲除细胞株进行验证重组RabMAb

重组Anti-CEACAM1抗体[EPR4049] (ab108397)

Submit a review Q&A (1)References (8)
Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 antibody [EPR4049] (ab108397)
  • Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
  • Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
  • Immunoprecipitation - Anti-CEACAM1 antibody [EPR4049] (ab108397)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 antibody [EPR4049] (ab108397)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-CEACAM1抗体[EPR4049]
    参阅全部 CEACAM1 一抗

  • 描述

    兔单克隆抗体[EPR4049] to CEACAM1

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IP, IHC-Pmore details
    不适用于: Flow Cyt

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Human fetal liver, A549, SW480, and HepG2 cell lysates; IHC-P: Human kidney clear cell carcinoma tissue. IP: SW480 cells

  • 常规说明

    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
    For more information see here.

    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.

性能

  • 形式

    Liquid

  • 存放说明

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.

  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • 纯度

    Protein A purified

  • 克隆

    单克隆

  • 克隆编号

    EPR4049

  • 同种型

    IgG

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab108397于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
1/10000 - 1/50000. Predicted molecular weight: 58 kDa.
IP
1/10 - 1/100.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
说明
WB
1/10000 - 1/50000. Predicted molecular weight: 58 kDa.
IP
1/10 - 1/100.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
应用说明
Is unsuitable for Flow Cyt.

靶标

  • 序列相似性

    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.

  • 细胞定位

    Secreted; Cell membrane and Cell membrane. Localizes to sites of cell-cell contact.
  • Information by UniProt

  • 数据库链接

    • 形式

      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.

    • 别名

      • Antigen CD66 antibody
      • BGP 1 antibody
      • BGP antibody
      • BGP-1 antibody
      • BGPI antibody
      • Biliary glycoprotein 1 antibody
      • Biliary glycoprotein adhesion molecule antibody
      • Biliary glycoprotein antibody
      • Carcinoembryonic antigen related cell adhesion molecule 1 antibody
      • carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein) antibody
      • Carcinoembryonic antigen-related cell adhesion molecule 1 antibody
      • CD66a antibody
      • CD66a antigen antibody
      • CEACAM1 antibody
      • CEAM1_HUMAN antibody
      • meconium antigen 100 antibody
      see all

    图片

    • Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      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.

    • Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      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.

    • Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      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

    • Immunoprecipitation - Anti-CEACAM1 antibody [EPR4049] (ab108397)
      Immunoprecipitation - Anti-CEACAM1 antibody [EPR4049] (ab108397)

      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)
      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.

    • Anti-CEACAM1 antibody [EPR4049] (ab108397)
      Anti-CEACAM1 antibody [EPR4049] (ab108397)

    文献 (8)

    发表研究结果有使用 ab108397？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab108397 被引用在 8 文献中.

    • Wu J  et al. Exosomes in malignant pleural effusion from lung cancer patients impaired the cytotoxicity of double-negative T cells. Transl Oncol 27:101564 (2023). PubMed: 36252282
    • Liu D  et al. Transfer RNA-derived fragment 5'tRF-Gly promotes the development of hepatocellular carcinoma by direct targeting of carcinoembryonic antigen-related cell adhesion molecule 1. Cancer Sci 113:3476-3488 (2022). PubMed: 35879647
    • Mattox AK  et al. Myeloid Cells Are Enriched in Tonsillar Crypts, Providing Insight into the Viral Tropism of Human Papillomavirus. Am J Pathol 191:1774-1786 (2021). PubMed: 34303699
    • Chean J  et al. Human CEACAM1-LF regulates lipid storage in HepG2 cells via fatty acid transporter CD36. J Biol Chem 297:101311 (2021). PubMed: 34666041
    • Suzuki S  et al. Immune-checkpoint molecules on regulatory T-cells as a potential therapeutic target in head and neck squamous cell cancers. Cancer Sci 111:1943-1957 (2020). PubMed: 32304268
    • Xu D  et al. PD-L1 Expression Is Regulated By NF-?B During EMT Signaling In Gastric Carcinoma. Onco Targets Ther 12:10099-10105 (2019). PubMed: 31819504
    • Wei Y  et al. DcR3 promotes proliferation and invasion of pancreatic cancer via a DcR3/STAT1/IRF1 feedback loop. Am J Cancer Res 9:2618-2633 (2019). PubMed: 31911850
    • Ye S  et al. Deep sequencing of primary human lung epithelial cells challenged with H5N1 influenza virus reveals a proviral role for CEACAM1. Sci Rep 8:15468 (2018). PubMed: 30341336

    客户评价及客户问答

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    Question

    Hello, I am searching a CEACAM antibody that can detect CEACAM on MDCK cells, which come from dog kidney. I am wondering if by immunogen peptied homology comparison it is possible to find one antibody that is suitable for dog CEACAM1 or other more common CEACAMs. Would some anti-human pan CEACAM work on dog CEACAMs? I would very much appreciate it if you can help me pick up the right antibody.

    Read More
    Answer

    Thank you for your inquiry. I was liaising with the laboratories to find out more about our pan-CEACAM- and CEACAM1-antibodies. Unfortunately, it turned out that we do not have any suitable antibody available: The immunogens of our CEACAM1 antibodies share a sequence homology with dog of 67% at the best. Thus, we would not predict a cross-reactivity for one of these antibodies. The same applies for our pan-CEACAM antibodies: The human CEACAM proteins used as immunogens reveal sequence similarities with the dog ortholog that are below 70%, suggesting that a cross-reactivity of the antibody is rather unlikely. If you have not done yet, I would like to recommend checking the Biocompare website which has an excellent antibody search facility that includes many suppliers. The links are: www.biocompare.com http://www.biocompare.com/ProductCategories/2045/Antibodies.html I am sorry that I could not be more helpful on this occasion and hope this information is nevertheless helpful to you. Please do not hesitate to contact me if you have any further questions in this regard.

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