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PRODUCT CODE: EM1901-75

Cytokeratin 19 Mouse Monoclonal Antibody [A3D1] (EM1901-75)

  • IVD–IHC

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

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All lanes: Western blot analysis of Cytokeratin 19 with anti-Cytokeratin 19 antibody [A3D1] (EM1901-75) at 1:1,000 dilution.<br /> Lane 1/2: Wild-type MCF-7 whole cell lysate (20 µg).<br /> Lane 3/4: Cytokeratin 19 fragment 1 knockdown MCF-7 whole cell lysate (20 µg).<br /> Lane 5/6: Cytokeratin 19 fragment 2 knockdown MCF-7 whole cell lysate (20 µg).<br /> <br /> EM1901-75 was shown to specifically react with Cytokeratin 19 in wild-type MCF-7 cells. Weakened bands were observed when Cytokeratin 19 knockdown samples were tested. Wild-type and Cytokeratin 19 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (EM1901-75, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
  • All lanes: Western blot analysis of Cytokeratin 19 with anti-Cytokeratin 19 antibody [A3D1] (EM1901-75) at 1:1,000 dilution.<br /> Lane 1/2: Wild-type MCF-7 whole cell lysate (20 µg).<br /> Lane 3/4: Cytokeratin 19 fragment 1 knockdown MCF-7 whole cell lysate (20 µg).<br /> Lane 5/6: Cytokeratin 19 fragment 2 knockdown MCF-7 whole cell lysate (20 µg).<br /> <br /> EM1901-75 was shown to specifically react with Cytokeratin 19 in wild-type MCF-7 cells. Weakened bands were observed when Cytokeratin 19 knockdown samples were tested. Wild-type and Cytokeratin 19 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (EM1901-75, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Cytokeratin 19 on MCF-7 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-75, 1/5000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-Cytokeratin 19 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 (EM1901-75, 1/1000) 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 small intestine tissue using anti-Cytokeratin 19 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 (EM1901-75, 1/1000) 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 pancreas tissue using anti-Cytokeratin 19 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 (EM1901-75, 1/1000) 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 colon cancer tissue using anti-Cytokeratin 19 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 (EM1901-75, 1/1000) 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 19 was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-75, 1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa FluorTM488 Goat anti-Mouse IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
All lanes: Western blot analysis of Cytokeratin 19 with anti-Cytokeratin 19 antibody [A3D1] (EM1901-75) at 1:1,000 dilution.
Lane 1/2: Wild-type MCF-7 whole cell lysate (20 µg).
Lane 3/4: Cytokeratin 19 fragment 1 knockdown MCF-7 whole cell lysate (20 µg).
Lane 5/6: Cytokeratin 19 fragment 2 knockdown MCF-7 whole cell lysate (20 µg).

EM1901-75 was shown to specifically react with Cytokeratin 19 in wild-type MCF-7 cells. Weakened bands were observed when Cytokeratin 19 knockdown samples were tested. Wild-type and Cytokeratin 19 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (EM1901-75, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

Cytokeratin 19 Mouse Monoclonal Antibody [A3D1] (EM1901-75)

Immunogen

Recombinant protein within human keratin 19 aa 128-322.

Host

Mouse

Positive Control

MCF-7 cell lysates, human breast tissue, human small intestine tissue, human pancreas tissue, human colon cancer tissue, MCF-7.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

A3D1

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

44 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:2000-1:10000

  • IHC-P

  • 1:500-1:2000

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

P08727

PROTEIN NAME

Cytokeratin 19

SYNONYMS

40 kDa keratin intermediate filament antibody; CK 19 antibody; CK-19 antibody; CK19 antibody; Cytokeratin 19 antibody; Cytokeratin-19 antibody; K19 antibody; K1C19_HUMAN antibody; K1CS antibody; Keratin 19 antibody; Keratin type I 40 kD antibody; Keratin type I 40kD antibody; Keratin type I cytoskeletal 19 antibody; Keratin, type I cytoskeletal 19 antibody; Keratin, type I, 40 kd antibody; Keratin-19 antibody; KRT19 antibody; MGC15366 antibody

SEQUENCE SIMILARITIES

Belongs to the intermediate filament family.

TISSUE SPECIFICITY

Expressed in a defined zone of basal keratinocytes in the deep outer root sheath of hair follicles. Also observed in sweat gland and mammary gland ductal and secretory cells, bile ducts, gastrointestinal tract, bladder urothelium, oral epithelia, esophagus, ectocervical epithelium (at protein level). Expressed in epidermal basal cells, in nipple epidermis and a defined region of the hair follicle. Also seen in a subset of vascular wall cells in both the veins and artery of human umbilical cord, and in umbilical cord vascular smooth muscle. Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma in structures that contain dystrophin and spectrin.

DEVELOPMENTAL STAGE

Present in hair follicles at all stages of development.

SUBCELLULAR LOCATION

Cytoskeleton, Cytosol.

FUNCTION

The keratin gene family is a large group of intermediate filament and structural proteins that provide the structural components of hair, nails, horns, scales, claws and similar types of hard tissues. Keratins are also important for intracellular stability in epithelial tissues and different keratins often display organ- or tissue specific expression patterns. Due to these specific expression patterns, keratins are often used as diagnostic biomarkers. Keratin 19 is a type I keratin and expressed in simple epithelia including glandular cell types present in the GI-tract, female tissues, male tissues, and respiratory epithelium. Clinically, keratin 19 is used together with keratin 18 to distinguish hepatocellular cancer from cholangiocellular carcinoma (both keratins are expressed in bile ducts, but keratin 18 is expressed in hepatocytes whereas keratin 19 is not). Keratin 19 is a type I keratin. The type I cytokeratins consist of acidic proteins which are arranged in pairs of heterotypic keratin chains. Unlike its related family members, this smallest known acidic cytokeratin is not paired with a basic cytokeratin in epithelial cells. It is specifically found in the periderm, the transiently superficial layer that envelops the developing epidermis. The type I cytokeratins are clustered in a region of chromosome 17q12-q21.