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Cytokeratin 7 Recombinant Mouse Monoclonal Antibody [3-G3-D8-R]

Usd: 350

Specification

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Catalog# HA601065

Cytokeratin 7 Recombinant Mouse Monoclonal Antibody [3-G3-D8-R]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

  • IF-Tissue

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

Cytokeratin 7 Recombinant Mouse Monoclonal Antibody [3-G3-D8-R]

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human Cytokeratin 7.

Species Reactivity

Human

Validated Applications

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

Target Molecular Weight

Predicted band size: 51 kDa

Positive Control

SK-OV-3 (Human ovarian cancer cell) cell lysate, HeLa (Human cervical adenocarcinoma cell) cell lysate, human breast tissue, human lung cancer tissue, HeLa, A549.

Conjugation

unconjugated

Clone Number

3-G3-D8-R

RRID

AB_3071788

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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

  • 1:1,000-1:5,000

  • IHC-P

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:100

  • FC

  • 1:500-1:1,000

  • IF-Tissue

  • 1:1,000

Target

Function

Keratin-7 is a member of the keratin gene family. The type II cytokeratins consist of basic or neutral proteins which are arranged in pairs of heterotypic keratin chains coexpressed during differentiation of simple and stratified epithelial tissues. Keratin-7 is found in simple glandular epithelia, and in transitional epithelium. Epithelial cells of the lung and breast both contain keratin-7, but some other glandular epithelia, such as those of the colon and prostate, do not. Because the keratin-7 antigen is found in both healthy and neoplastic cells, antibodies to CK7 can be used in immunohistochemistry to distinguish ovarian and transitional cell carcinomas from colonic and prostate cancers, respectively. Pancreas is recommended as positive tissue control for CK7. Virtually all ductal and intercalated duct epithelial cells must show an at least weak to moderate, distinct cytoplasmic staining reaction, whereas epithelial cells of large ducts should display a strong staining intensity. Acinar epithelial cells should be negative. In general, no staining reaction should be seen in the epithelial cells. Scattered columnar epithelial cells and endothelial cells may show a weak to moderate cytoplasmic staining reaction.

Background References

1. Lin L., Holbro T., Alonso G., Gerosa D., Burger M.M. J. Molecular interaction between human tumor marker protein p150, the largest subunit of eIF3, and intermediate filament protein K7. Cell. Biochem. 80:483-490(2001)

2. Smith F.J.D., Porter R.M., Corden L.D., Lunny D.P., Lane E.B., McLean W.H.I. Cloning of human, murine, and marsupial keratin 7 and a survey of K7 expression in the mouse. Biochem. Biophys. Res. Commun. 297:818-827(2002)

3. Sanden C., Broselid S., Cornmark L., Andersson K., Daszkiewicz-Nilsson J., Martensson U.E., Olde B., Leeb-Lundberg L.M. G protein-coupled estrogen receptor 1/G protein-coupled receptor 30 localizes in the plasma membrane and traffics intracellularly on cytokeratin intermediate filaments. Mol. Pharmacol. 79:400-410(2011)

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: P08729 Human

Synonyms

CK 7 antibody

CK-7 antibody

CK7 antibody

Cytokeratin 7 antibody

Cytokeratin-7 antibody

D15Wsu77e antibody

K2C7 antibody

K2C7_HUMAN antibody

K7 antibody

Keratin 7 antibody

Expand

Images

  • Western blot analysis of Cytokeratin 7 on different lysates with Mouse anti-Cytokeratin 7 antibody (<a href="/products/HA601065" style="font-weight: bold;text-decoration: underline;">HA601065</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate<br />Lane 2: SK-OV-3 (Human ovarian cancer cell) cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 3 minutes; ECL: K1801<br /><br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA601065" style="font-weight: bold;text-decoration: underline;">HA601065</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Mouse IgG-HRP (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 51 kDa<br />Observed band size: 51 kDa

    Western blot analysis of Cytokeratin 7 on different lysates with Mouse anti-Cytokeratin 7 antibody (HA601065) at 1/5,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
    Lane 2: SK-OV-3 (Human ovarian cancer cell) cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 3 minutes; ECL: K1801


    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA601065, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 51 kDa
    Observed band size: 51 kDa

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Breast cancer<br />Sample: Paraffin-embedded section<br /><br />Primary antibody dilution: 1/500<br />Antigen retrieval: ER2<br />Platform: Leica Biosystems BOND&reg; RX

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Breast cancer
    Sample: Paraffin-embedded section

    Primary antibody dilution: 1/500
    Antigen retrieval: ER2
    Platform: Leica Biosystems BOND® RX

  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Mouse anti-Cytokeratin 7 antibody (<a href="/products/HA601065" style="font-weight: bold;text-decoration: underline;">HA601065</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/HA601065" style="font-weight: bold;text-decoration: underline;">HA601065</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 cancer tissue with Mouse anti-Cytokeratin 7 antibody (HA601065) 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 (HA601065) 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.

  • Immunocytochemistry analysis of A549 cells labeling Cytokeratin 7 with Mouse anti-Cytokeratin 7 antibody (<a href="/products/HA601065" style="font-weight: bold;text-decoration: underline;">HA601065</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 7 antibody (<a href="/products/HA601065" style="font-weight: bold;text-decoration: underline;">HA601065</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 A549 cells labeling Cytokeratin 7 with Mouse anti-Cytokeratin 7 antibody (HA601065) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 7 antibody (HA601065) 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.

  • Flow cytometric analysis of HeLa cells labeling Cytokeratin 7.<br /><br />Cells were fixed and permeabilized, and then blocked with 2% negative goat serum for 15 minutes at room temperature.Then stained with the primary antibody (<a href="/products/HA601065" style="font-weight: bold;text-decoration: underline;">HA601065</a>, 1ug/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 7.

    Cells were fixed and permeabilized, and then blocked with 2% negative goat serum for 15 minutes at room temperature.Then stained with the primary antibody (HA601065, 1ug/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).

  • Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: lung carcinoma<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/1,000

    Application: IF-Tissue

    Species: Human

    Site: lung carcinoma

    Sample: Paraffin-embedded section

    Antibody concentration: 1/1,000

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