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Cytokeratin 9 Recombinant Rabbit Monoclonal Antibody [SD201-09]

Usd: 385

Specification

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Catalog# ET1612-77

Cytokeratin 9 Recombinant Rabbit Monoclonal Antibody [SD201-09]

Application

  • WB

  • IHC-P

  • IF-Tissue

  • IF-Cell

  • IP

Reactivity

  • Human

  • Mouse

BSA and Azide free
Predicted reactivity

  • Rat

Predicted species support after-sales service
Conjugation

  • unconjugated

Overview

Product Name

Cytokeratin 9 Recombinant Rabbit Monoclonal Antibody [SD201-09]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein wiehin Human Cytokeratin 9 aa 234-422 / 623.

Species Reactivity

Human, Mouse (Predicted: Rat)

Predicted species support after-sales service

Validated Applications

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

Molecular Weight

Predicted band size: 62 kDa

Positive Control

HAP-1 cell lysates, mouse skin tissue, human breast tissue, MCF-7.

Conjugation

unconjugated

Clone Number

SD201-09

RRID

AB_3070153

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

  • IHC-P

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IF-Cell

  • 1:50

Target

Function

Cytokeratins comprise a diverse group of intermediate filament proteins that are expressed as pairs in both keratinized and non-keratinized epithelial tissue. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. Cytokeratin 9 is an unusually large, type I acidic cytokeratin that differentiates human plantar and palmar epidermal cells. Cytokeratin 9 localizes to the suprabasal layers as well as the upper epidermal layers such as the glandular ridges and interridges. The domains of cytokeratin 9 include a head, an α-helical coiled-coil-forming rod, and a tail; cytokeratin 9 shares significant homology with cytokeratin 10. Mutations in the cytokeratin 9 gene correlate with the development of epidermolytic palmoplantar keratoderma (EPPK), an autosomal dominant inherited skin disorder that is characterized by hyperkeratosis of the skin over the palms and soles.

Background References

1. De Rosa, L. et al. 2014. Long-term stability and safety of transgenic cultured epidermal stem cells in gene therapy of junctional epidermolysis bullosa. Stem Cell Reports. 2: 1-8.

2. Rice RH et al. 2013. Distinguishing ichthyoses by protein profiling. PLoS One 8:e75355.

Sequence Similarity

Belongs to the intermediate filament family.

Tissue Specificity

Expressed in the terminally differentiated epidermis of palms and soles.

Subcellular Location

Extracellular exosome, membrane, nucleus.

UNIPROT

SwissProt: P35527 Human

SwissProt: Q6RHW0 Mouse

SwissProt: Q8CIS9 Rat

Synonyms

CK 9 antibody

CK9 antibody

CK-9 antibody

Cytokeratin 9 antibody

Cytokeratin-9 antibody

Cytokeratin9 antibody

EPPK antibody

K1C9_HUMAN antibody

K9 antibody

Keratin 9 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Cytokeratin 9 on different lysates with Rabbit anti-Cytokeratin 9 antibody (<a href="/products/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Cytokeratin 9 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 62 kDa<br />Observed band size: 62 kDa<br /><br />Exposure time: 18 seconds; ECL: K1801;<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/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Cytokeratin 9 on different lysates with Rabbit anti-Cytokeratin 9 antibody (ET1612-77) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Cytokeratin 9 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 62 kDa
    Observed band size: 62 kDa

    Exposure time: 18 seconds; ECL: K1801;

    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 (ET1612-77) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-Cytokeratin 9 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% 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/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</a>, 1/50) 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 mouse skin tissue using anti-Cytokeratin 9 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-77, 1/50) 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.

  • Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 9 (<a href="/products/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</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 9 (<a href="/products/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</a>, red) at 1/50 dilution and Vimentin (<a href="/products/EM0401" style="font-weight: bold;text-decoration: underline;">EM0401</a>, green) at 1/500 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 breast tissue labeling Cytokeratin 9 (ET1612-77) 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 9 (ET1612-77, red) at 1/50 dilution and Vimentin (EM0401, green) at 1/500 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).

  • Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 9 (<a href="/products/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</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 9 (<a href="/products/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</a>, red) at 1/50 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>) was used as the secondary antibodies at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 9 (ET1612-77).

    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 9 (ET1612-77, red) at 1/50 dilution at +4℃ overnight, washed with PBS.

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

  • Immunocytochemistry analysis of MCF-7 cells labeling Cytokeratin 9 (<a href="/products/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</a>).<br /><br />Cells were fixed in 4% paraformaldehyde and permeabilized with 0.05% Triton X-100 in PBS for 10 minutes, and then blocked with 2% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody Cytokeratin 9 (<a href="/products/ET1612-77" style="font-weight: bold;text-decoration: underline;">ET1612-77</a>, red) at 1/200 dilution for overnight at 4℃.<br /><br />Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) was used as the secondary antibody at 1/800 dilution. DAPI was used to stain the cell nuclei (blue).

    Immunocytochemistry analysis of MCF-7 cells labeling Cytokeratin 9 (ET1612-77).

    Cells were fixed in 4% paraformaldehyde and permeabilized with 0.05% Triton X-100 in PBS for 10 minutes, and then blocked with 2% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody Cytokeratin 9 (ET1612-77, red) at 1/200 dilution for overnight at 4℃.

    Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/800 dilution. DAPI was used to stain the cell nuclei (blue).

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