Structured Review

Agilent technologies proteinase k
Ubiquitin and α-syn aggregates colocalize with neuronal markers. Sections of hippocampus in 12-mo-old Gba1 D409V/D409V brains, either pretreated with <t>proteinase</t> K (PK) ( B ) or not ( A and C ) before immunostaining. Immunostaining for NFH ( A ) exposed
Proteinase K, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 716 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/proteinase k/product/Agilent technologies
Average 94 stars, based on 716 article reviews
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94/100 stars

Images

1) Product Images from "CNS expression of glucocerebrosidase corrects ?-synuclein pathology and memory in a mouse model of Gaucher-related synucleinopathy"

Article Title: CNS expression of glucocerebrosidase corrects ?-synuclein pathology and memory in a mouse model of Gaucher-related synucleinopathy

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1108197108

Ubiquitin and α-syn aggregates colocalize with neuronal markers. Sections of hippocampus in 12-mo-old Gba1 D409V/D409V brains, either pretreated with proteinase K (PK) ( B ) or not ( A and C ) before immunostaining. Immunostaining for NFH ( A ) exposed
Figure Legend Snippet: Ubiquitin and α-syn aggregates colocalize with neuronal markers. Sections of hippocampus in 12-mo-old Gba1 D409V/D409V brains, either pretreated with proteinase K (PK) ( B ) or not ( A and C ) before immunostaining. Immunostaining for NFH ( A ) exposed

Techniques Used: Immunostaining

Progressive accumulation of α-syn/ubiquitin aggregates in Gba1 D409V/D409V Gaucher mouse brains. ( A ) Frozen sections were pretreated without ( Upper ) or with ( Lower ) proteinase K (PK) to uncover endogenous α-syn aggregates (arrowheads).
Figure Legend Snippet: Progressive accumulation of α-syn/ubiquitin aggregates in Gba1 D409V/D409V Gaucher mouse brains. ( A ) Frozen sections were pretreated without ( Upper ) or with ( Lower ) proteinase K (PK) to uncover endogenous α-syn aggregates (arrowheads).

Techniques Used:

2) Product Images from "Analytic Variability in Immunohistochemistry Biomarker Studies"

Article Title: Analytic Variability in Immunohistochemistry Biomarker Studies

Journal: Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology

doi: 10.1158/1055-9965.EPI-10-0097

Representative AQUA output for 31G7 after progressive proteinase K digestion for 5, 10, 15, and 45 min at room temperature. Analysis of pan-cytokeratin used as a control for tumor integrity revealed similar results.
Figure Legend Snippet: Representative AQUA output for 31G7 after progressive proteinase K digestion for 5, 10, 15, and 45 min at room temperature. Analysis of pan-cytokeratin used as a control for tumor integrity revealed similar results.

Techniques Used:

3) Product Images from "Abnormal Neurites Containing C-Terminally Truncated ?-Synuclein Are Present in Alzheimer's Disease without Conventional Lewy Body Pathology"

Article Title: Abnormal Neurites Containing C-Terminally Truncated ?-Synuclein Are Present in Alzheimer's Disease without Conventional Lewy Body Pathology

Journal: The American Journal of Pathology

doi: 10.2353/ajpath.2010.100552

Truncated α-syn is detected by immunohistochemistry. Lewy neurites and Lewy bodies containing C-terminally truncated α-syn forms α-syn110 and α-syn119 are present in LBV ( A , B , D , and E ), and Lewy neurites containing α-syn119 are present in AD ( G ). Proteinase K pretreatment enhances the immunohistochemical signal for α-syn110 and α-syn119 in Lewy neurites ( B , E , G ) compared with sections not pretreated with proteinase K ( A , D , F ), which suggests that the detected truncated α-syn is in an aggregated form. Proteinase K pretreatment was necessary for the detection of any α-syn119-positive neurites in AD ( F , G ). Preabsorption of the antibody with the respective antigenic peptide abolishes the signal ( C , H ), which supports the specificity of the signal. (Immunohistochemistry, cingulate cortex; red chromogen; Scale bar = 20 μm).
Figure Legend Snippet: Truncated α-syn is detected by immunohistochemistry. Lewy neurites and Lewy bodies containing C-terminally truncated α-syn forms α-syn110 and α-syn119 are present in LBV ( A , B , D , and E ), and Lewy neurites containing α-syn119 are present in AD ( G ). Proteinase K pretreatment enhances the immunohistochemical signal for α-syn110 and α-syn119 in Lewy neurites ( B , E , G ) compared with sections not pretreated with proteinase K ( A , D , F ), which suggests that the detected truncated α-syn is in an aggregated form. Proteinase K pretreatment was necessary for the detection of any α-syn119-positive neurites in AD ( F , G ). Preabsorption of the antibody with the respective antigenic peptide abolishes the signal ( C , H ), which supports the specificity of the signal. (Immunohistochemistry, cingulate cortex; red chromogen; Scale bar = 20 μm).

Techniques Used: Immunohistochemistry

4) Product Images from "Neonatal AAV delivery of alpha-synuclein induces pathology in the adult mouse brain"

Article Title: Neonatal AAV delivery of alpha-synuclein induces pathology in the adult mouse brain

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-017-0455-3

Detection of αsyn-associated pathology in AAV1-αsyn mouse. a , b Photomicrographs of representative regions of the brain of AAV1-αsyn injected mice.at 3 months of age. a Phosphorylated αsyn (pS129) was highly increased within the neuronal soma and to a lesser extent within the axonal projections.5G4 immunostaining was less intense but follow the same pattern as pS129. Neither pS129 nor 5G4 were found in AAV-venus animals ( bottom line ). b Brain sections digested with proteinase K showed PK-resistant αsyn in neuronal cell bodies and neurites with small inclusions (
Figure Legend Snippet: Detection of αsyn-associated pathology in AAV1-αsyn mouse. a , b Photomicrographs of representative regions of the brain of AAV1-αsyn injected mice.at 3 months of age. a Phosphorylated αsyn (pS129) was highly increased within the neuronal soma and to a lesser extent within the axonal projections.5G4 immunostaining was less intense but follow the same pattern as pS129. Neither pS129 nor 5G4 were found in AAV-venus animals ( bottom line ). b Brain sections digested with proteinase K showed PK-resistant αsyn in neuronal cell bodies and neurites with small inclusions (

Techniques Used: Injection, Mouse Assay, Immunostaining

5) Product Images from "Characterization of Expression of Glycan Ligands for Siglec-F in Normal Mouse Lungs"

Article Title: Characterization of Expression of Glycan Ligands for Siglec-F in Normal Mouse Lungs

Journal: American Journal of Respiratory Cell and Molecular Biology

doi: 10.1165/rcmb.2010-0007OC

Binding of Siglec-F–Ig fusion protein to normal mouse lung without ( A ) or with ( B ) proteinase K treatment. Staining in both models was completely eliminated by protease treatment. Results are shown from one of three experiments with similar results.
Figure Legend Snippet: Binding of Siglec-F–Ig fusion protein to normal mouse lung without ( A ) or with ( B ) proteinase K treatment. Staining in both models was completely eliminated by protease treatment. Results are shown from one of three experiments with similar results.

Techniques Used: Binding Assay, Staining

6) Product Images from "Evaluation of the Possible Transmission of BSE and Scrapie to Gilthead Sea Bream (Sparus aurata)"

Article Title: Evaluation of the Possible Transmission of BSE and Scrapie to Gilthead Sea Bream (Sparus aurata)

Journal: PLoS ONE

doi: 10.1371/journal.pone.0006175

Progressive increase in the size of proteinase K resistant deposits in BSE-challenged fish. The mean diameter of the immunohistochemically detected (SaurPrP1) deposits after proteinase K–digestion is given with reference to time p.i..
Figure Legend Snippet: Progressive increase in the size of proteinase K resistant deposits in BSE-challenged fish. The mean diameter of the immunohistochemically detected (SaurPrP1) deposits after proteinase K–digestion is given with reference to time p.i..

Techniques Used: Fluorescence In Situ Hybridization

Related Articles

Clone Assay:

Article Title: Analytic Variability in Immunohistochemistry Biomarker Studies
Article Snippet: .. Sections (5 μm thick) were deparaffinized, rehydrated, and antigen retrieved by pressure cooking in 10 mmol/L citrate (pH 6) or 10 mmol/L Tris/1 mmol/L EDTA buffer (pH 9) for all primary antibodies except anti-EGFR clones 2-18C9 (pharmDx; Dako, Carpinteria, CA) and 31G7 (Zymed/Invitrogen, Carlsbad, CA), for which proteinase K (Dako) digestion was done instead. .. Slides were incubated with a cocktail of the primary antibody and a mouse monoclonal cytokeratin (clone AE1/AE3, M3515; Dako) or a polyclonal rabbit cytokeratin antibody (Z0622; Dako) overnight at 4°C for all antibodies except ER clone 1D5 and EGFR clone 2-18C9, which were incubated for 1 h at room temperature ( ).

Immunohistochemistry:

Article Title: Abnormal Neurites Containing C-Terminally Truncated ?-Synuclein Are Present in Alzheimer's Disease without Conventional Lewy Body Pathology
Article Snippet: .. For immunohistochemistry with the syn110, syn119, syn122-140 (LB509), and N-ter syn antibodies , unless specified otherwise, the sections were pretreated with proteinase K (#53020; Dako, Carpinteria, CA) for 15 minutes before incubation in a 1:200, 1:200, 1:80, or 1:100 dilution of the primary antibody, respectively, for 30 minutes. at room temperature (RT). .. For the negative control experiment with a preabsorbed primary antibody, an aliquot of the primary antibody was incubated with a 100-fold molar excess of the antigenic peptide resuspended in distilled water with 2 mmol/L dithiothreitol for 30 minutes with gentle shaking at RT.

Avidin-Biotin Assay:

Article Title: A biosafety level-2 dose-dependent lethal mouse model of spotted fever rickettsiosis: Rickettsia parkeri Atlantic Rainforest strain
Article Snippet: .. After that, they were blocked with Avidin/Biotin Blocking Kit (Life Technologies, Frederick, MD) and treated with proteinase K (Dako, Carpinteria, CA), for antigen retrieval. .. The sections were incubated with polyclonal rabbit anti-R . conorii antibody (1:300 dilution, produced in-house) at room temperature for one hour, followed by biotinylated secondary anti-rabbit IgG (1:200 dilution, Vector Laboratories, Burlingame, CA), streptavidin-AP (1:200, Vector Laboratories, Burlingame, CA) for 30 minutes each and Fast Red (Dako, Carpinteria, CA) for 5 minutes.

Immunostaining:

Article Title: Neonatal AAV delivery of alpha-synuclein induces pathology in the adult mouse brain
Article Snippet: .. Lastly, for proteinase K (PK) digestion, tissue sections were preincubated with proteinase K (DAKO) in PBS for 2 min at room temperature before performing regular immunostaining for αsyn using LB509 antibody (Thermo fisher). .. Western blot Frozen hemi brains were mechanically homogenized on ice in 10% (w /v ) of cold lysis RIPA buffer (Millipore) containing protease inhibitor cocktail (Roche Diagnostics), and centrifuged at 100,000 x g for 20 min.

Incubation:

Article Title: Combining radiation with autophagy inhibition enhances suppression of tumor growth and angiogenesis in esophageal cancer
Article Snippet: .. Briefly, following deparaffinization and dehydration, the tissue sections were incubated in proteinase K (DAKO North America, Inc., Carpinteria, CA, USA) for 15 min, washed with PBS, incubated in equilibration buffer and then in terminal deoxynucleotidyl transferase enzyme solution. .. The sections were subsequently rinsed in PBS, incubated with streptavidin-peroxidase conjugate (Sigma-Aldrich) and visualized using diaminobenzidine (Sigma-Aldrich), according to the manufacturer’s instructions.

Article Title: Abnormal Neurites Containing C-Terminally Truncated ?-Synuclein Are Present in Alzheimer's Disease without Conventional Lewy Body Pathology
Article Snippet: .. For immunohistochemistry with the syn110, syn119, syn122-140 (LB509), and N-ter syn antibodies , unless specified otherwise, the sections were pretreated with proteinase K (#53020; Dako, Carpinteria, CA) for 15 minutes before incubation in a 1:200, 1:200, 1:80, or 1:100 dilution of the primary antibody, respectively, for 30 minutes. at room temperature (RT). .. For the negative control experiment with a preabsorbed primary antibody, an aliquot of the primary antibody was incubated with a 100-fold molar excess of the antigenic peptide resuspended in distilled water with 2 mmol/L dithiothreitol for 30 minutes with gentle shaking at RT.

Article Title: Evaluation of the Possible Transmission of BSE and Scrapie to Gilthead Sea Bream (Sparus aurata)
Article Snippet: .. For PrPSc detection, the sections were hydrated-autoclaved at 121°C for 30 minutes, then incubated for 5 minutes in 90% formic acid prior to an 8 minutes-incubation with proteinase K (Dako, Glostrup, Denmark) at RT. .. Sections were treated with appropriate biotinylated secondary antibodies (Vector Laboratories, Burlingame, CA) and visualized using the avidin-biotin method-based Vectastain Elite ABC and the Diaminobezidine substrate kits (Vector Laboratories, Burlingame, CA) according to the manufacturer's instructions.

Blocking Assay:

Article Title: A biosafety level-2 dose-dependent lethal mouse model of spotted fever rickettsiosis: Rickettsia parkeri Atlantic Rainforest strain
Article Snippet: .. After that, they were blocked with Avidin/Biotin Blocking Kit (Life Technologies, Frederick, MD) and treated with proteinase K (Dako, Carpinteria, CA), for antigen retrieval. .. The sections were incubated with polyclonal rabbit anti-R . conorii antibody (1:300 dilution, produced in-house) at room temperature for one hour, followed by biotinylated secondary anti-rabbit IgG (1:200 dilution, Vector Laboratories, Burlingame, CA), streptavidin-AP (1:200, Vector Laboratories, Burlingame, CA) for 30 minutes each and Fast Red (Dako, Carpinteria, CA) for 5 minutes.

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    Agilent technologies proteinase k buffer
    Proteinase K Buffer, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/proteinase k buffer/product/Agilent technologies
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    proteinase k buffer - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    94
    Agilent technologies proteinase k
    Proteinase K, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 716 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/proteinase k/product/Agilent technologies
    Average 94 stars, based on 716 article reviews
    Price from $9.99 to $1999.99
    proteinase k - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

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