Anti-Cytokeratin 8+18 antibody [EP1628Y] (ab53280)

Overview

  • Product nameAnti-Cytokeratin 8+18 antibody [EP1628Y]
    See all Cytokeratin 8+18 primary antibodies
  • Description
    Rabbit monoclonal [EP1628Y] to Cytokeratin 8+18
  • SpecificityThis ab also cross react with Cytokeratin 18
  • Tested applicationsSuitable for: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Cytokeratin 8+18 aa 300-400 (C terminal). The exact sequence is proprietary.
    Database link: P05787

  • Positive control
    • A431 cell lysate, HeLa cells or human breast adenocarcinoma tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

Properties

Applications

Our Abpromise guarantee covers the use of ab53280 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
WB 1/25000 - 1/50000. Detects a band of approximately 52 kDa (predicted molecular weight: 54 kDa).
IP 1/70.
Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration.
ICC/IF 1/100 - 1/500.

Target

  • RelevanceFunction: Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle. Tissue specificity: Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma membrane in structures that contain dystrophin and spectrin. Expressed in gingival mucosa and hard palate of the oral cavity. Disease: Cirrhosis Similarity: Belongs to the intermediate filament family. PTM: Phosphorylation on serine residues is enhanced during EGF stimulation and mitosis. Ser-74 phosphorylation plays an important role in keratin filament reorganization. O-glycosylated. O-GlcNAcylation at multiple sites increases solubility, and decreases stability by inducing proteasomal degradation. O-glycosylated (O-GlcNAcylated), in a cell cycle-dependent manner.
  • Cellular localizationCytoplasmic
  • Database links
  • Alternative names
    • CARD2 antibody
    • Cell proliferation inducing gene 46 protein antibody
    • Cell proliferation inducing protein 46 antibody
    • CK 8 antibody
    • CK-8 antibody
    • CK18 antibody
    • CK8 antibody
    • CYK18 antibody
    • CYK8 antibody
    • Cytokeratin 18 antibody
    • Cytokeratin 8 antibody
    • Cytokeratin-8 antibody
    • K18 antibody
    • K2C8 antibody
    • K2C8_HUMAN antibody
    • K8 antibody
    • Keratin 18 antibody
    • Keratin 8 antibody
    • Keratin antibody
    • Keratin type I cytoskeletal 18 antibody
    • keratin type II cytoskeletal 8 antibody
    • Keratin-8 antibody
    • KO antibody
    • KRT18 antibody
    • KRT8 antibody
    • type II cytoskeletal 8 antibody
    • Type-II keratin Kb8 antibody
    see all

Anti-Cytokeratin 8+18 antibody [EP1628Y] images

  • Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

    Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)

    Preparation:

    Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 µm thickness

    Primary antibody 1: Rabbit anti cytokeratin 8 (ab53280), 1:100

    Primary antibody 2: Rat anti-perlecan, 1:100
    Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200

    Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed (ab150081), 1:200
    Nuclei were counterstained with DAPI

     

  • ab53280 staining Cytokeratin 8+18 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Cytokeratin 8+18 with purified ab53280 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised by 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

  • Capture ab: 1/20 dilution (ab133272, 2μg in 0.35mg lysates) 

    Primary ab for WB:
    - Panel A: CK-18 (ab133272,1/1000)
    - Panel B: CK-8 (ab53280,1/1000)

    Secondary ab: VeriBlot for IP secondary antibody (HRP) (ab131366)
    Blocking and dilution buffer: 5% NFDM/TBST

    Lane 1 (Input): HeLa (human cervix adenocarcinoma) whole cell lysate 10ug
    Lane 2 (+): HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133272 in HeLa (human cervix adenocarcinoma) whole cell lysate

    Exposure time:
    - Panel A: 30 seconds
    - Panel B: 1 seconds

     

  • Ab53280 (1:250) staining human Cyotkeratin 8 in human breast adenocarcinoma tissue by immunohistochemistry using paraffin embedded tissue.Ab53280 (1:250) staining human Cyotkeratin 8 in human breast adenocarcinoma tissue by immunohistochemistry using paraffin embedded tissue.
  • Anti-Cytokeratin 8+18 antibody [EP1628Y] (ab53280) at 1/50000 dilution + A431 cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labeled at 1/2000 dilution

    Predicted band size : 54 kDa
    Observed band size : 52 kDa (why is the actual band size different from the predicted?)
  • ab53280 staining Cytokeratin 8 in Mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formalin and blocked with 10% serum for 20 minutes at 23°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/75 in TBS + 1% BSA) for 1 hour at 23°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunofluorescent staining of HeLa cells using ab53280 (1:100).
  • Fluorescent immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue using ab53280. Green-CK8 red-PI.

  • Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab53280. Green-CK8 red-PI

  • Fluorescent immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue using ab53280. Green-CK8 red-PI.

  • Fluorescent immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue using ab53280. Green-CK8 red-PI.

  • Overlay histogram showing HeLa cells stained with ab53280 (red line). The cells were fixed with 2% PFA (room temperature, 30 min) and then permeabilized with 1% FACS permeabilizing solution for 30 min. The cells were then incubated in 3% FBS in 1X PBS followed by the antibody (ab53280, 1/20 dilution) for 1 hour at room temperature. The cells were then incubated for 30 min at room temperature with the secondary antibody. An isotype control antibody (black line) was used and an unlabelled sample (blue line) was also used as a control.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-Cytokeratin 8+18 antibody [EP1628Y] (ab53280)

This product has been referenced in:
  • Ruiz A  et al. Effect of hydroxychloroquine and characterization of autophagy in a mouse model of endometriosis. Cell Death Dis 7:e2059 (2016). IHC-P ; Mouse . Read more (PubMed: 26775710) »
  • Hubbs AF  et al. Accumulation of Ubiquitin and Sequestosome-1 Implicate Protein Damage in Diacetyl-Induced Cytotoxicity. Am J Pathol N/A:N/A (2016). ICC/IF ; Mouse . Read more (PubMed: 27643531) »

See all 14 Publications for this product

Product Wall

The immunogen sequence that generated cytokeratin antibody ab53280 is 93% identical to the rat cytokeratin type II cytoskeletal 8, accession NP_955402.1. So we expect cross-reaction but do not have experimental evidence. The potential for cross-reactio...

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Bos taurus Tissue sections (Oviduct, epithelial cells)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.0
Permeabilization No
Specification Oviduct, epithelial cells
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Nov 27 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample Mouse Cell (Differentiated P19 cell line)
Specification Differentiated P19 cell line
Permeabilization Yes - Triton X-100
Fixative Formaldehyde
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Submitted Jul 24 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Liver)
Specification Liver
Fixative Formalin
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 23°C
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Verified customer

Submitted Mar 04 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"