Recombinant
RabMAb

Recombinant Anti-Cytokeratin 18 antibody [SP69] - BSA and Azide free (ab198380)

Overview

  • Product name

    Anti-Cytokeratin 18 antibody [SP69] - BSA and Azide free
    See all Cytokeratin 18 primary antibodies
  • Description

    Rabbit monoclonal [SP69] to Cytokeratin 18 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Cytokeratin 18 aa 400 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P05783

  • Positive control

    • IHC-P: Human prostate or breast carcinoma, Human stomach, and Human endometrium tissue; FC: HeLa cells; ICC: HeLa cells.
  • General notes

    Ab198380 is the carrier-free version of ab93741. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab198380 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab198380 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 Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Incubate for 30 minutes at RT. Perform heat mediated antigen retrieval before commencing with IHC staining protocol by boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.

Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Involved in the uptake of thrombin-antithrombin complexes by hepatic cells (By similarity). When phosphorylated, plays a role in filament reorganization. Involved in the delivery of mutated CFTR to the plasma membrane. Together with KRT8, is involved in interleukin-6 (IL-6)-mediated barrier protection.
  • Tissue specificity

    Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.
  • Involvement in disease

    Defects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600].
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Post-translational
    modifications

    Phosphorylation at Ser-34 increases during mitosis. Hyperphosphorylated at Ser-53 in diseased cirrhosis liver. Phosphorylation increases by IL-6.
    Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-238 by either caspase-3, caspase-6 or caspase-7.
    O-glycosylated at multiple sites; glycans consist of single N-acetylglucosamine residues.
  • Cellular localization

    Cytoplasm > perinuclear region.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cell proliferation inducing gene 46 protein antibody
    • Cell proliferation inducing protein 46 antibody
    • Cell proliferation-inducing gene 46 protein antibody
    • CK 18 antibody
    • CK-18 antibody
    • CK18 antibody
    • CYK 18 antibody
    • CYK18 antibody
    • Cytokeratin 18 antibody
    • Cytokeratin endo B antibody
    • Cytokeratin-18 antibody
    • K 18 antibody
    • K18 antibody
    • K1C18_HUMAN antibody
    • KA18 antibody
    • Keratin 18 antibody
    • Keratin 18, type I antibody
    • Keratin D antibody
    • keratin, type I cytoskeletal 18 antibody
    • Keratin-18 antibody
    • Krt18 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrium tissue sections labeling Cytokeratin 18 with ab93741 at 1/200 dilution (1.21 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab93741)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach tissue sections labeling Cytokeratin 18 with ab93741 at 1/200 dilution (1.21 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab93741)
  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab93741 at 1:100(2.4 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab135384).
  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab93741 at 1:240 dilution (1.00 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab135384).
  • ICC/IF image of ab93741 stained HCT116 cells.

    The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab93741 at 1/500 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti-Rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab93741).

  • Flow cytometric analysis of rabbit anti-Cytokeratin 18 (SP69) antibody ab93741(1/100) in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (green) compared to negative control of rabbit IgG (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab93741).

  • ab93741 at 1/200 dilution staining Cytokeratin 18 in formalin-fixed, paraffin-embedded human breast carcinoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab93741).

  • Overlay histogram showing MCF7 (Human breast adenocarcinoma cell line) cells stained with ab93741 (red line).

    The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab93741, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabeled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab93741).

  • Immunihistochemical staining of human stomach with ab93741.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab93741).

References

ab198380 has not yet been referenced specifically in any publications.

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