Cytokeratin 19 Antibody - BSA Free
Novus Biologicals | Catalog # NBP1-78278
![Western Blot: Cytokeratin 19 AntibodyBSA Free [NBP1-78278]](https://resources.rndsystems.com/images/products/Cytokeratin-19-Antibody-Western-Blot-NBP1-78278-img0006.jpg "Western Blot: Cytokeratin 19 AntibodyBSA Free [NBP1-78278]")
Key Product Details
Species Reactivity
Validated:
Human, Rat
Cited:
Rat, Ovine
Applications
Validated:
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western
Cited:
Immunohistochemistry-Paraffin, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Format
BSA Free
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Product Specifications
Immunogen
A synthetic peptide made to an N-terminal portion of the human Cytokeratin 19 protein (between residues 1-50) [UniProt P08727]
Reactivity Notes
Rat reactivity reported in scientific literature (PMID: 27070090).
Localization
Cytoplasmic.
Marker
Epithelial Cell Marker
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Cytokeratin 19 Antibody - BSA Free
Western Blot: Cytokeratin 19 AntibodyBSA Free [NBP1-78278]
Western Blot: Cytokeratin 19 Antibody [NBP1-78278] - Analysis of Cytokeratin 19 in MCF7 lysate.![Immunocytochemistry/ Immunofluorescence: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]](https://resources.rndsystems.com/images/products/Cytokeratin-19-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-78278-img0004.jpg "Immunocytochemistry/ Immunofluorescence: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]")
Immunocytochemistry/ Immunofluorescence: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]
Immunocytochemistry/Immunofluorescence: Cytokeratin 19 Antibody [NBP1-78278] - Antibody at 1:25 in MCF7 cells with FITC (green). Nuclei were counterstained with DAPI (blue).![Immunohistochemistry: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]](https://resources.rndsystems.com/images/products/Cytokeratin-19-Antibody-Immunohistochemistry-NBP1-78278-img0005.jpg "Immunohistochemistry: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]")
Immunohistochemistry: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]
Immunohistochemistry: Cytokeratin 19 Antibody [NBP1-78278] - Analysis of Cytokeratin 19 in human kidney carcinoma using DAB with hematoxylin counterstain.![Immunocytochemistry/ Immunofluorescence: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]](https://resources.rndsystems.com/images/products/Cytokeratin-19-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-78278-img0007.jpg "Immunocytochemistry/ Immunofluorescence: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]")
Immunocytochemistry/ Immunofluorescence: Cytokeratin 19 Antibody - BSA Free [NBP1-78278]
Immunocytochemistry/Immunofluorescence: Cytokeratin 19 Antibody [NBP1-78278] - Confocal immunofluorescent analysis of HepG2 cells using Cytokeratin 19 antibody (NBP1-78278, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).![Simple Western: Cytokeratin 19 AntibodyBSA Free [NBP1-78278]](https://resources.rndsystems.com/images/products/Cytokeratin-19-Antibody-Simple-Western-NBP1-78278-img0008.jpg "Simple Western: Cytokeratin 19 AntibodyBSA Free [NBP1-78278]")
Simple Western: Cytokeratin 19 AntibodyBSA Free [NBP1-78278]
Simple Western: Cytokeratin 19 Antibody [NBP1-78278] - Image shows a specific band for Cytokeratin 19 in 0.05 mg/mL of MCF-7 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.Applications for Cytokeratin 19 Antibody - BSA Free
Application
Recommended Usage
Immunocytochemistry/ Immunofluorescence
1:75
Immunohistochemistry
1:400
Immunohistochemistry-Paraffin
1:400
Simple Western
1:3000
Western Blot
2 ug/mL
Application Notes
In Western blot a band is seen at ~44 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in MCF-7 lysate 0.05 mg/mL, separated by Size, antibody dilution of 1:3000, apparent MW was 51 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in MCF-7 lysate 0.05 mg/mL, separated by Size, antibody dilution of 1:3000, apparent MW was 51 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
Reviewed Applications
Read 2 reviews rated 4.5 using NBP1-78278 in the following applications:
Formulation, Preparation, and Storage
Purification
Immunogen affinity purified
Formulation
PBS, 30% Glycerol
Format
BSA Free
Preservative
0.05% Sodium Azide
Concentration
1.16 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: Cytokeratin 19
Additional Cytokeratin 19 Products
Product Documents for Cytokeratin 19 Antibody - BSA Free
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Product Specific Notices for Cytokeratin 19 Antibody - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Related Research Areas
Citations for Cytokeratin 19 Antibody - BSA Free
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Customer Reviews for Cytokeratin 19 Antibody - BSA Free (2)
4.5 out of 5
2 Customer Ratings
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Application: ImmunocytochemistrySample Tested:Species: HumanVerified Customer | Posted 06/09/2014Confocal immunofluorescent analysis of HepG2 cells using Cytokeratin 19 antibody (NBP1-78278, 1:5).
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Application: Western BlotSample Tested:Species: HumanVerified Customer | Posted 06/09/2014Western blot analysis of extracts from MCF7 and HepG2 cells using Cytokeratin 19 antibody (NBP1-78278, 1:100).
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Protocols
View specific protocols for Cytokeratin 19 Antibody - BSA Free (NBP1-78278):
Cytokeratin 19 Antibody:
Immunocytochemistry Protocol
Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.
1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Immunocytochemistry Protocol
Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.
1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Cytokeratin 19 Antibody:
Immunohistochemistry-Paraffin Embedded Sections
Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.
Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Immunohistochemistry-Paraffin Embedded Sections
Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.
Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Cytokeratin 19 Antibody:
Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Cytokeratin 19 Antibody - BSA Free
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Q: Would you be able to confirm whether or not NBP1-78278 - Cytokeratin 19 Antibody is mouse reactive?
A: We have not yet tested our product NBP1-78278 with mouse samples. Based on an alignment of the immunogen used to raise this product and the mouse sequence, this product shares 71% homology with the mouse protein.
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Associated Pathways
