Overview

  • Product name
    Anti-Cytokeratin 19 antibody [BA-17]
    See all Cytokeratin 19 primary antibodies
  • Description
    Mouse monoclonal [BA-17] to Cytokeratin 19
  • Host species
    Mouse
  • Specificity
    Cytokeratin peptide 19 (40 kDa) in human tissue.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-Fr, WB, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Mammary organoids.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    >95% by SDS-PAGE
  • Clonality
    Monoclonal
  • Clone number
    BA-17
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7755 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. PubMed: 16870717
WB Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

Target

  • Function
    Involved in the organization of myofibers. Together with KRT8, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle.
  • 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.
  • Sequence similarities
    Belongs to the intermediate filament family.
  • Developmental stage
    Present in hair follicles at all stages of development.
  • Domain
    This keratin differs from all other IF proteins in lacking the C-terminal tail domain.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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
    see all

Images

  • All lanes : Anti-Cytokeratin 19 antibody [BA-17] (ab7755) at 5 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Observed band size: 44 kDa
    why is the actual band size different from the predicted?

  • Cytokeratin 19 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Mouse monoclonal to Cytokeratin 19 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab7755.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 44kDa: Cytokeratin 19
  • Human normal skin. Staining is observed in the cytoplasm (epidermal basal cells). Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for mouse for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ICC/IF image of ab7755 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7755, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Anti-Cytokeratin 19 antibody [BA-17] (ab7755) + Cell lysates prepared from human MCF-7 cells
  • Overlay histogram showing MCF7 cells stained with ab7755 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab7755, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Zhao Z  et al. Intraductal papillary mucinous neoplasm of the pancreas rapidly xenografts in chicken eggs and predicts aggressiveness. Int J Cancer 142:1440-1452 (2018). IHC-Fr ; Human . Read more (PubMed: 29143337) »
  • Yamazaki M  et al. Transcriptional regulation of human amelotin gene by interleukin-1ß. FEBS Open Bio 8:974-985 (2018). Read more (PubMed: 29928577) »
See all 11 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 1mM EDTA in 10mM Tris-HCl Buffer (pH=9.0)
Permeabilization
No
Specification
Liver
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 28 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (Colon cancer cell lines)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Colon cancer cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Jun 05 2015

Answer

Thank you for contacting us.

You had previously been in contact with my colleague heather regarding staining observed in your negative control using these mouse monoclonal Cytokeratin antibodies on mouse tissue. To clarify, is your negative control from a tissue that is not expected to express cytokeratin or is it an isotype control or a no primary antibody control?

When using an anti-mouse secondary antibody on mouse tissue, background staining is often observed due to cross-reactivity with endogenous mouse IgGs in the sample. This can be checked by testing a no-primary antibody control slide.

If you are observing mouse on mouse background staining, this can be avoided by either using a mouse on mouse (MOM) staining kit, such as ab127055. The background can also be blocked by adding an incubation with a F(ab) fragment unconjugated anti-mouse secondary antibody, such as ab6668. The F(ab) fragment will coat any endogenous IgGs in your sample, and can be done just before the serum block at a 1:500 dilution for 1 hour at room temperature.

I hope this helps, if not, please let me know and I will be happy to help you further.

Read More

Question
Answer

I am very pleased to hear you would like to accept our offer and test ab7755 in rat. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for rat and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Human pancreatic neoplastic lesions)
Specification
Human pancreatic neoplastic lesions
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
No
Blocking step
MOM kit form VectorLab as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 12 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (pancreatic intraepithelial neoplasia (PanINs))
Specification
pancreatic intraepithelial neoplasia (PanINs)
Fixative
acetone+methanol 1:1
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 06 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (pancreatic cancer)
Loading amount
35 µg
Specification
pancreatic cancer
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Dr. Pawel Mazur

Verified customer

Submitted Jun 09 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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