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Cytokeratin 4 Recombinant Rabbit Monoclonal Antibody [SN74-03]

Usd: 385

Specification

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Catalog# ET1611-71

Cytokeratin 4 Recombinant Rabbit Monoclonal Antibody [SN74-03]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

Cytokeratin 4 Recombinant Rabbit Monoclonal Antibody [SN74-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Cytokeratin 4 aa 395-520 / 520.

Species Reactivity

Human

Validated Applications

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

Molecular Weight

Predicted band size: 56 kDa

Positive Control

A431, human tonsil tissue, human esophagus tissue.

Conjugation

unconjugated

Clone Number

SN74-03

RRID

AB_3070051

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:1,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:100-1:500

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

Target

Function

Cytokeratins are a subfamily of intermediate filament keratins that are characterized by a remarkable biochemical diversity, which is represented in human epithelial tissues by at least 20 different polypeptides. Cytokeratins range in isoelectric range between 4.9 and 7.8. Cytokeratin 1 has the highest molecular weight, while Cytokeratin 19 has the lowest molecular weight. The cytokeratins are divided into the type I and type II subgroups. Type II family members comprise the basic to neutral members, Cytokeratins 1-8, while the type I group comprises the acidic members, Cytokeratins 9-20. Various epithelia in the human body usually express cytokeratins which are characteristic of the type of epithelium and related to the degree of maturation or differentiation within said epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. Cytokeratin 4 is expressed in differentiated layers of the mucosal and esophageal epithelia along with Cytokeratin 13.

Background References

1. Jacot TA et al. Development of a composite measure of product adherence, protocol compliance, and semen exposure using DNA and protein biomarkers for topical HIV prevention studies. PLoS One 9:e114368 (2014).

2. Badylak SF et al. Esophageal preservation in five male patients after endoscopic inner-layer circumferential resection in the setting of superficial cancer: a regenerative medicine approach with a biologic scaffold. Tissue Eng Part A 17:1643-50 (2011).

Sequence Similarity

Belongs to the intermediate filament family.

Tissue Specificity

Detected in the suprabasal layer of the stratified epithelium of the esophagus, exocervix, vagina, mouth and lingual mucosa, and in cells and cell clusters in the mucosa and serous gland ducts of the esophageal submucosa (at protein level). Expressed widely in the exocervix and esophageal epithelium, with lowest levels detected in the basal cell layer.

Subcellular Location

Cell surface, intermediate filament cytoskeleton, nucleus.

UNIPROT

SwissProt: P19013 Human

Synonyms

CK 4 antibody

CK-4 antibody

CK4 antibody

CYK4 antibody

Cytokeratin 4 antibody

Cytokeratin-4 antibody

Cytokeratin4 antibody

FLJ31692 antibody

K2C4_HUMAN antibody

K4 antibody

Expand

Images

  • Immunocytochemistry analysis of A431 cells labeling Cytokeratin 4 with Rabbit anti-Cytokeratin 4 antibody (<a href="/products/ET1611-71" style="font-weight: bold;text-decoration: underline;">ET1611-71</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 Rabbit anti-Cytokeratin 4 antibody (<a href="/products/ET1611-71" style="font-weight: bold;text-decoration: underline;">ET1611-71</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of A431 cells labeling Cytokeratin 4 with Rabbit anti-Cytokeratin 4 antibody (ET1611-71) 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 Rabbit anti-Cytokeratin 4 antibody (ET1611-71) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling Cytokeratin 4 (<a href="/products/ET1611-71" style="font-weight: bold;text-decoration: underline;">ET1611-71</a>) and Vimentin (<a href="/products/EM0401" style="font-weight: bold;text-decoration: underline;">EM0401</a>).<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 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Cytokeratin 4 (<a href="/products/ET1611-71" style="font-weight: bold;text-decoration: underline;">ET1611-71</a>, red) at 1/200 dilution and Vimentin (<a href="/products/EM0401" style="font-weight: bold;text-decoration: underline;">EM0401</a>, green) at 1/200 dilution at +4℃ overnight, washed with PBS.<br /><br />Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) and Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) were used as the secondary antibodies at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling Cytokeratin 4 (ET1611-71) and Vimentin (EM0401).

    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 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Cytokeratin 4 (ET1611-71, red) at 1/200 dilution and Vimentin (EM0401, green) at 1/200 dilution at +4℃ overnight, washed with PBS.

    Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) and Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) were used as the secondary antibodies at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human esophagus tissue using anti-Cytokeratin 4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-71" style="font-weight: bold;text-decoration: underline;">ET1611-71</a>, 1/400) for 30 minutes 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 using anti-Cytokeratin 4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-71, 1/400) for 30 minutes 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 using anti-Cytokeratin 4 antibody.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-71" style="font-weight: bold;text-decoration: underline;">ET1611-71</a>, 1/400) for 30 minutes 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 using anti-Cytokeratin 4 antibody.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-71, 1/400) for 30 minutes 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 Cytokeratin 4 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1611-71" style="font-weight: bold;text-decoration: underline;">ET1611-71</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Cytokeratin 4 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-71, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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