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Cytokeratin 18 Mouse Monoclonal Antibody [6-19]

Usd: 350

Specification

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Catalog# M0407-19

Cytokeratin 18 Mouse Monoclonal Antibody [6-19]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Cytokeratin 18 Mouse Monoclonal Antibody [6-19]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within mouse Cytokeratin 18 aa 374-423 / 423.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 48 kDa

Positive Control

HeLa cell lysate, K-562 cell lysate, A431 cell lysate, HT-29 cell lysate, HepG2 cell lysate, HCT 116 cell lysate, Huh7 cell lysate, HeLa, human breast cancer tissue, human liver tissue, mouse liver tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

6-19

RRID

AB_3072981

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG2a

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:10,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:2,000

  • FC

  • 1:1,000

Target

Function

Cytokeratin 18 is an acidic keratin which is found primarily in non squamous epithelia and is present in a majority of adenocarcinomas and ductal carcinomas but not in squamous cell carcinomas. Cytokeratin 18 exists in combination with Cytokeratin 8, a basic keratin. Hepatocellular carcinomas have been reportedly defined by the use of antibodies that recognize only Cytokeratins 8 and 18.

Background References

1. Korver S et al. The application of cytokeratin-18 as a biomarker for drug-induced liver injury. Arch Toxicol. 2021 Nov

2. Goralska J et al. Plasma Cytokeratin-18 Fragment Level Reflects the Metabolic Phenotype in Obesity. Biomolecules. 2023 Apr

Sequence Similarity

Belongs to the intermediate filament family.

Tissue Specificity

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

Post-translational Modification

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-GlcNAcylation increases solubility, and decreases stability by inducing proteasomal degradation.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P05783 Human

SwissProt: P05784 Mouse

SwissProt: Q5BJY9 Rat

Synonyms

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

Expand

Images

  • Western blot analysis of Cytokeratin 18 on different lysates with Mouse anti-Cytokeratin 18 antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: K-562 cell lysate<br />Lane 3: A431 cell lysate<br />Lane 4: HT-29 cell lysate<br />Lane 5: HepG2 cell lysate<br />Lane 6: HCT 116 cell lysate<br />Lane 7: Huh7 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 48 kDa<br />Observed band size: 48 kDa<br /><br />Exposure time: 5 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Cytokeratin 18 on different lysates with Mouse anti-Cytokeratin 18 antibody (M0407-19) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: K-562 cell lysate
    Lane 3: A431 cell lysate
    Lane 4: HT-29 cell lysate
    Lane 5: HepG2 cell lysate
    Lane 6: HCT 116 cell lysate
    Lane 7: Huh7 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 48 kDa
    Observed band size: 48 kDa

    Exposure time: 5 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M0407-19) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling Cytokeratin 18 with Mouse anti-Cytokeratin 18 antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Cytokeratin 18 antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Cytokeratin 18 with Mouse anti-Cytokeratin 18 antibody (M0407-19) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Cytokeratin 18 antibody (M0407-19) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Mouse anti-Cytokeratin 18 antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Mouse anti-Cytokeratin 18 antibody (M0407-19) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M0407-19) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-Cytokeratin 18 antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-Cytokeratin 18 antibody (M0407-19) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M0407-19) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-Cytokeratin 18 antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-Cytokeratin 18 antibody (M0407-19) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M0407-19) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue with Mouse anti-Cytokeratin 18 antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue with Mouse anti-Cytokeratin 18 antibody (M0407-19) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M0407-19) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling Cytokeratin 18.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/M0407-19" style="font-weight: bold;text-decoration: underline;">M0407-19</a>, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Cytokeratin 18.

    Cells were fixed and permeabilized. Then stained with the primary antibody (M0407-19, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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