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Cytokeratin 14 Recombinant Mouse Monoclonal Antibody [A2C10-R]

Usd: 350

Specification

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Catalog# HA601323

Cytokeratin 14 Recombinant Mouse Monoclonal Antibody [A2C10-R]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Cytokeratin 14 Recombinant Mouse Monoclonal Antibody [A2C10-R]

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within Human Cytokeratin 14 aa 423-472 / 472.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IF-Tissue

Molecular Weight

Predicted band size: 52 kDa

Positive Control

A431 cell lysate, mouse skin tissue lysate, rat skin tissue lysate, A431, human breast tissue, human esophagus tissue, human lung squamous carcinoma tissue, human tonsil tissue, mouse esophagus tissue.

Conjugation

unconjugated

Clone Number

A2C10-R

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000

  • IF-Tissue

  • 1:200

Target

Function

This gene encodes a member of the keratin family, the most diverse group of intermediate filaments. This gene product, a type I keratin, is usually found as a heterotetramer with two keratin 5 molecules, a type II keratin. Together they form the cytoskeleton of epithelial cells. Mutations in the genes for these keratins are associated with epidermolysis bullosa simplex. The nonhelical tail domain is involved in promoting KRT5-KRT14 filaments to self-organize into large bundles and enhances the mechanical properties involved in resilience of keratin intermediate filaments in vitro. Expressed in the corneal epithelium (at protein level). Detected in the basal layer, lowered within the more apically located layers specifically in the stratum spinosum, stratum granulosum but is not detected in stratum corneum. Strongly expressed in the outer root sheath of anagen follicles but not in the germinative matrix, inner root sheath or hair. A form of epidermolysis bullosa simplex, a group of skin fragility disorders characterized by skin blistering due to cleavage within the basal layer of keratinocytes, and erosions caused by minor mechanical trauma. There is a broad spectrum of clinical severity ranging from minor blistering on the feet, to subtypes with extracutaneous involvement and a lethal outcome. EBS1A is an autosomal dominant form characterized by generalized intraepidermal skin blistering that begins and is very prominent at birth. EBS1A may be life-threatening in the first year of life. Tendency to blistering diminishes in adolescence.

Background References

1. Bousquet O. et al. The nonhelical tail domain of keratin 14 promotes filament bundling and enhances the mechanical properties of keratin intermediate filaments in vitro. J. Cell Biol. 155:747-754(2001).

2. Schweizer J et al. "New consensus nomenclature for mammalian keratins". The Journal of Cell Biology. 174 (2): 169–74(2006).

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P02533 Human

SwissProt: Q61781 Mouse

SwissProt: Q6IFV1 Rat

Synonyms

CK 14 antibody

CK-14 antibody

ck14 antibody

Cytokeratin 14 antibody

Cytokeratin-14 antibody

Cytokeratin14 antibody

Dowling Meara antibody

EBS3 antibody

EBS4 antibody

Epidermolysis bullosa simplex antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Cytokeratin 14 on different lysates with Mouse anti-Cytokeratin 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate (20 µg/Lane)<br />Lane 2: PANC-1 cell lysate (negative) (20 µg/Lane)<br />Lane 3: Mouse skin tissue lysate (40 µg/Lane)<br />Lane 4: Rat skin tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 1 minute;<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/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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.

    ☑ Relative expression (RE)

    Western blot analysis of Cytokeratin 14 on different lysates with Mouse anti-Cytokeratin 14 antibody (HA601323) at 1/1,000 dilution.

    Lane 1: A431 cell lysate (20 µg/Lane)
    Lane 2: PANC-1 cell lysate (negative) (20 µg/Lane)
    Lane 3: Mouse skin tissue lysate (40 µg/Lane)
    Lane 4: Rat skin tissue lysate (40 µg/Lane)

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: 1 minute;

    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 (HA601323) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of A431 (positive) and PANC-1 (negative) labeling Cytokeratin 14 with Mouse anti-Cytokeratin 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</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 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</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.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of A431 (positive) and PANC-1 (negative) labeling Cytokeratin 14 with Mouse anti-Cytokeratin 14 antibody (HA601323) 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 14 antibody (HA601323) 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 tissue with Mouse anti-Cytokeratin 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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 tissue with Mouse anti-Cytokeratin 14 antibody (HA601323) at 1/1,000 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 (HA601323) at 1/1,000 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 esophagus tissue with Mouse anti-Cytokeratin 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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 esophagus tissue with Mouse anti-Cytokeratin 14 antibody (HA601323) at 1/1,000 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 (HA601323) at 1/1,000 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 lung squamous carcinoma tissue with Mouse anti-Cytokeratin 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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 lung squamous carcinoma tissue with Mouse anti-Cytokeratin 14 antibody (HA601323) at 1/1,000 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 (HA601323) at 1/1,000 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 tonsil tissue with Mouse anti-Cytokeratin 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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 tonsil tissue with Mouse anti-Cytokeratin 14 antibody (HA601323) at 1/1,000 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 (HA601323) at 1/1,000 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 esophagus tissue with Mouse anti-Cytokeratin 14 antibody (<a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>) at 1/1,000 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 esophagus tissue with Mouse anti-Cytokeratin 14 antibody (HA601323) at 1/1,000 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 (HA601323) at 1/1,000 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.

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Mouse<br />Tissue: Skin<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>, 1/200, overnight at 4℃.<br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Mouse
    Tissue: Skin
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
    Primary antibody: HA601323, 1/200, overnight at 4℃.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Rat<br />Tissue: Skin<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601323" style="font-weight: bold;text-decoration: underline;">HA601323</a>, 1/200, overnight at 4℃.<br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Rat
    Tissue: Skin
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
    Primary antibody: HA601323, 1/200, overnight at 4℃.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

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