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Signal Transduction Cytoskeleton / ECM Cytoskeleton Intermediate Filaments Class I Cytokeratins
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Recombinant

Recombinant Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

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Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
  • Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [C-11] to pan Cytokeratin - BSA and Azide free
  • Suitable for: IHC-P, IHC-Fr, ICC/IF, mIHC
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Unconjugated

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Anti-pan Cytokeratin antibody [KRT/1877R] (ab234297)
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Anti-wide spectrum Cytokeratin antibody (ab9377)

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Overview

  • Product name

    Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free
    See all pan Cytokeratin primary antibodies
  • Description

    Mouse monoclonal [C-11] to pan Cytokeratin - BSA and Azide free
  • Host species

    Mouse
  • Specificity

    Cytokeratin peptides 4,5,6,8,10,13,18.

  • Tested applications

    Suitable for: IHC-P, IHC-Fr, ICC/IF, mIHCmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Tissue, cells or virus. This information is considered to be commercially sensitive.

  • Positive control

    • IHC-Fr: Rat kidney, mouse large intestine and human skin. IHC-P: Rat, mouse and human skin. ICC/IF: HeLa and A431 cells. mIHC: Human tonsil tissue and Human breast cancer tissue.
  • General notes

    ab264485 is the carrier-free version of ab7753.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    Purified from TCS. Purity >95% by SDS-PAGE.
  • Clonality

    Monoclonal
  • Clone number

    C-11
  • Isotype

    IgG1
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class I
    • Cytokeratins

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-pan Cytokeratin antibody [C-11] (ab277270)
    • Anti-pan Cytokeratin antibody [C-11] (ab7753)
  • Recombinant Protein

    • Anti-pan Cytokeratin antibody [KRT/1877R] (ab234297)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab264485 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
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
mIHC
Use at an assay dependent concentration.
Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
mIHC
Use at an assay dependent concentration.

Target

  • Relevance

    Cytokeratins, a group comprising at least 29 different proteins, are characteristic of epithelial and trichocytic cells. Cytokeratins 1, 4, 5, 6, and 8 are members of the type II neutral to basic subfamily. Monoclonal anti cytokeratins are specific markers of epithelial cell differentiation and have been widely used as tools in tumor identification and classification. Monoclonal Anti Pan Cytokeratin is a broadly reactive reagent, which recognizes epitopes present in most human epithelial tissues. It facilitates typing of normal, metaplastic and neoplastic cells. Synergy between the various components results in staining amplification. This enables identification of cells, which would otherwise be stained only marginally. The mixture may aid in the discrimination of carcinomas and nonepithelial tumors such as sarcomas, lymphomas and neural tumors. It is also useful in detecting micrometastases in lymph nodes, bone marrow and other tissues and for determining the origin of poorly differentiated tumors. There are two types of cytokeratins the acidic type I cytokeratins and the basic or neutral type II cytokeratins. Cytokeratins are usually found in pairs comprising a type I cytokeratin and a type II cytokeratin. Usually the type II cytokeratins are 8kD larger than their type I counterparts.
  • Cellular localization

    Cytoplasmic
  • Database links

    • Entrez Gene: 374454 Human
    • Entrez Gene: 51350 Human
    • SwissProt: Q01546 Human
    • SwissProt: Q7Z794 Human
    • Alternative names

      • pan ck antibody
      • pan-ck antibody
      • panck antibody

    Images

    • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

      IHC image of pan cytokeratin staining in a section of formalin-fixed paraffin-embedded normal human skin* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6,epitope retrieval solution 1) for 20mins. The section was then incubated with ab7753, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjµgated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions,primary antibody concentration and antibody incubation times.
      *Tissue obtained from the Human Research Tissue Bank,supported by the NIHR Cambridge Biomedical Research Centre

    • Immunocytochemistry/ Immunofluorescence - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunocytochemistry/ Immunofluorescence - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      ab7753 staining pan Cytokeratin in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min),permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab7753 at 1µg/ml and ab6046,Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117,Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080,Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min).

      Image was acquired with a high-content analyser (Operetta CLS,Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

       

      IHC image of pan cytokeratin staining in a section of frozen normal mouse large intestine performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab7753, 1µg/ml, for 15 mins at room temperature. A goat anti-mouse IgG1 bridging antibody,ab125913,was added for 8 mins at room temperature and detected using an HRP conjµgated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

       

      IHC image of pan cytokeratin staining in a section of frozen normal rat kidney performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab7753, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjµgated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    • Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

       

      IHC image of pan cytokeratin staining in a section of frozen normal human skin performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab7753, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjµgated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions,primary antibody concentration and antibody incubation times.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

       

      IHC image of pan cytokeratin staining in a section of formalin-fixed paraffin-embedded normal rat skin performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6,epitope retrieval solution 1) for 20mins. The section was then incubated with ab7753, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjµgated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions,primary antibody concentration and antibody incubation times.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

       

      IHC image of pan cytokeratin staining in a section of formalin-fixed paraffin-embedded normal mouse skin performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6,epitope retrieval solution 1) for 20mins. The section was then incubated with ab7753, 1µg/ml, for 15 mins at room temperature. A goat anti-mouse IgG1 bridging antibody,ab125913,was added for 8 mins at room temperature and detected using an HRP conjµgated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions,primary antibody concentration and antibody incubation times.

    • Immunocytochemistry/ Immunofluorescence - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunocytochemistry/ Immunofluorescence - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab7753)

      ab7753 staining pan Cytokeratin in A431 cells. The cells were fixed with 4% paraformaldehyde (10 min),permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab7753 at 1µg/ml and ab6046,Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117,Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080,Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min).

      Image was acquired with a high-content analyser (Operetta CLS,Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    • Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Multiplex immunohistochemistry - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

      This image was generated from the hybridoma version of the product.

       

      Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).

      Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).

       

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).

       

      The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.

       

      Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

       

      DAPI (dark blue) was used as a nuclear counter stain.

       

      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)This image is courtesy of an Abreview submitted by Carl Hobbs

      This image was generated from the hybridoma version of the product.

       

      ab7753 staining human skin sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab7753 at 1/250 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)This image is courtesy of an Abreview submitted by Carl Hobbs

      This image was generated from the hybridoma version of the product.

      ab7753 staining rat embryonic skin/organ sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab77539 at 1/250 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-mouse polyclonal antibody at 1/200 was used as the secondary antibody.

    • Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)
      Anti-pan Cytokeratin antibody [C-11] - BSA and Azide free (ab264485)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

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