We performed neuropathologic examinations on biopsied specimens taken from the white matter of anterior horn of lateral ventricle of Patient 5 and 7. Paraffin-embedded fixed tissue sections (7 μm) were stained with hematoxylin & eosin (H&E), and with periodic Schiff. Sections were also immunostained with a DAKO Autostainer system (DAKO, Carpinteria, CA) using EnVision FLEX DAB+ Chromogen with the following antibodies: anti-phosphorylated neurofilament (mouse monoclonal [2836], 1:100; Cell Signaling Technology); anti-CD68 (PGM1) (mouse, 1:100; Dako); anti-CD3 (D7A6E) (rabbit [85061], 1:200; Cell Signaling Technology); anti-CD20 (EP459Y) (rabbit [ab78237], 1:200; abcam); anti-Olig2 (EPR2673) (rabbit [ab109186], 1:100; abcam).
Dako autostainer system
Manufactured by Agilent Technologies
Sourced in Denmark
The DAKO Autostainer system is a fully automated immunohistochemistry (IHC) and in situ hybridization (ISH) staining platform. The system is designed to provide consistent and reliable staining results by automating the staining process, including reagent dispensing, incubation, and washing steps.
Automatically generated - may contain errors
Lab products found in correlation
Image pro plus 6, by Olympus (2 mentions)
Ab109186, by Abcam (1 mentions)
Anti cd68 pg m1, by Agilent Technologies (1 mentions)
Epr2673, by Abcam (1 mentions)
Envision flex detection system, by Agilent Technologies (1 mentions)
Dako autostainer link, by Agilent Technologies (1 mentions)
Ab76315, by Abcam (1 mentions)
Real detection system, by Agilent Technologies (1 mentions)
Bcl 2, by Abcam (1 mentions)
Lc3 1 2, by Proteintech (1 mentions)
K4003, by Agilent Technologies (1 mentions)
Envision flex hrp, by Agilent Technologies (1 mentions)
Envision flex mini kit, by Agilent Technologies (1 mentions)
Pd l1 ihc 22c3 pharmdx assay, by Agilent Technologies (1 mentions)
9 protocols using dako autostainer system
1
Neuropathological Examination of Anterior Horn
2
IHC Analysis of Xenograft Tumors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The IHC examination was performed on the tumors resected from our xenograft transplantation mice. The tissue samples were fixed with formalin, embedded in paraffin, and cut into 5-µm-thick sections for further Hematoxylin-eosin(H&E) staining or IHC staining. The DAKO Autostainer system (Dako, Glostrup, Denmark) was used for IHC staining. The sections were observed using an Olympus microscope (Tokyo, Japan), and the images were analyzed using Image-Pro Plus 6.0.
3
Immunohistochemical Analysis of pSTAT3 and Ki67
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hematoxylin and eosin (H&E) staining was performed by standard methods using a Dako Autostainer system (Dako, Agilent, Santa Clara, CA, USA). Immunohistochemistry was performed on formalin‐fixed, paraffin‐embedded tumour resection specimens. In brief, antigen retrieval was performed in Dako PT‐Link using EnVision™ FLEX Target Retrieval Solution (high pH). Immunohistochemical staining was performed in a Dako Autostainer Link using the EnVision™ FLEX detection system according to the manufacturer's instructions. Primary antibodies were anti phospo‐STAT3 (ab76315, Abcam) and anti Ki67 (MIB1, Dako). Sections of lobular breast carcinoma were used as a positive control, showing nuclear positivity in normal ductal structures as well as in scattered tumour cell nuclei. Processing of patient material and data has been approved by the Regional Research Ethics Committee (REC) in Gothenburg (reference number: 133‐11).
4
Colorectal Tumor Budding Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Colorectal specimens were macroscopically evaluated after fixing overnight in 10% buffered formalin. LNs were dissected using the methylene-blue method[10 (link),11 (link)]; samples from the resection margins, the tumor-region and other conspicuous areas were paraffin-embedded. The slides were stained with hematoxylin and eosin (HE) and evaluated by an experienced pathologist (BM). Based on the HE-morphology, slides were selected for further pan-cytokeratin staining which was performed to enable optimal evaluation of tumor budding. For this evaluation, monoclonal mouse antibody AE1/AE3 was used (dilution 1:50; DAKO). Immunoreactions were developed using a labelled streptavidin-biotin system (DAKO Real detection system). All reactions were performed on a Dako-Autostainer system (DAKO, Glostrup, Denmark).
Tumor budding was evaluated by one pathologist (BM). It was defined as detached single tumor cells or clusters of up to four cells. The cut-off for high-grade budding was adapted from Ueno et al[14 (link)] and defined as ≥ 30 buds/20 × magnification (= 1.3 mm²).
Tumor budding was evaluated by one pathologist (BM). It was defined as detached single tumor cells or clusters of up to four cells. The cut-off for high-grade budding was adapted from Ueno et al[14 (link)] and defined as ≥ 30 buds/20 × magnification (= 1.3 mm²).
5
Histological Analysis of Xenograft Tumors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue samples isolated from xenograft tumours were subjected to histological analysis. Briefly, tissue samples were first fixed with formalin and then embedded in paraffin, cut into 5 µm sections and stained with H&E for IHC staining. IHC staining was conducted in a DAKO Autostainer system (Dako, Glostrup, Denmark). The primary antibodies used were Bcl-2 (Abcam, ab32124), Ki-67 (Abcam, Cambridge, MA, UK), LC3-I/II (Proteintech, 14600-1-AP), p62 (MBL, M162-3). Primary tumours and major organ sections were stained with H&E. Images were visualised using an Olympus microscope (Japan), and image analysis was performed by Image-Pro Plus 6.0.
6
Immunohistochemical Analysis of NaPi-IIb
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After fixation, whole embryos were cut longitudinally and the two midsagittal halves were placed in a cassette and embedded in paraffin wax. Sections with 3–5 µm thickness were stained with hematoxylin and eosin (H&E) or subjected to immunohistochemical evaluation. Immunohistochemical analysis for NaPi-IIb was performed in the Dako autostainer system (Dako, Glostrup, Denmark) using the primary polyclonal antibody against rat NaPi-IIb indicated above. Briefly, upon antigen retrieval in citrate buffer (pH 6) at 98 °C for 20 min, sections were incubated overnight at 4 °C in the presence of the primary antibody (dilution 1:500). Upon application of EnVision-HRP anti-rabbit secondary antibody (Dako K4003) DAB substrate buffer (K3468) was used for the detection. As negative control, samples from wild type embryos were processed in parallel omitting the incubation with the primary antibody.
7
Immunohistochemical Analysis of WRAP53β and p53
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue microarrays were constructed as described previously.33 (link), 34 (link) Heat-mediated antigen retrieval (pH=9) was performed with the PT-link system and immunohistochemical staining in the DAKO Autostainer system (Dako, Glostrup, Denmark) using α-WRAP53 (1 : 25 dilution, # HPA023026, Atlas Antibodies, Stockholm, Sweden) and α-p53 (1 : 200 dilution, #AMAb90956, Atlas Antibodies). Normal matched fallopian tube samples with no evidence of histological disease (n=39) were stained as negative controls. Staining intensity of WRAP53β was assessed by two of the authors as 0=negative, 1=weak, 2=moderate or 3=strong. For statistical purposes, the staining scores were subdivided into low (0–2, n=139) and high (3, n=12). Staining intensity of p53 was also assessed by two of the authors as positive or negative.
8
Immunohistochemical Analysis of HLA Antigen Expression in Glioblastoma Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue specimens from resected glioblastoma tumors were analyzed by immunohistochemistry employing the EnVision FLEX Mini Kit and Dako Autostainer system (Agilent) following the manufacturer’s instructions. In brief, paraffin‐embedded tissues (3 μm) were deparaffinized and rehydrated and epitope retrieval was performed by heat induction (HIER) at 95°C for 20 min and pH = 9 in the PT Link, Pre‐Treatment Module. Endogenous peroxidase activity was blocked with EnVision FLEX Peroxidase‐Blocking Reagent for 5 min. Afterwards, sections were incubated with Protein Block, Serum‐Free buffer for 60 min. Tissue sections were stained with the following polyclonal antibodies: anti‐human HLA‐A (1:400, PA5‐116980), anti‐human HLA‐B (1:200, PA5‐35345), anti‐human HLA‐C (1:2000, PA5‐79367), anti‐human ILT2 (1:1000, PA5‐98738) (all from Invitrogen), or anti‐human HLA‐E (1:200, HPA031454, Merck) polyclonal antibodies and counterstained with hematoxylin. Signals were detected using diaminobenzidine chromogen as substrate in EnVision FLEX HRP (Agilent). Isotype controls were processed by omitting the primary antibody. Each specimen was individually reviewed and expression intensity was scored using a scale from 0 to 3. Sample classification was performed as follows: 0 as negative staining, 1 as dim, and 2–3 as positive.
9
PD-L1 IHC 22C3 Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PD-L1 expression was assessed in formalin-fixed tumor samples at a central laboratory with the use of the commercially available PD-L1 IHC 22C3 pharmDx assay (Dako North America; ref. 15) on the Dako Autostainer system (Agilent Technologies). Biopsies were obtained by core-needle. The scoring system used for PD-L1 expression was the Combined Positive Score 1 (CPS1), measuring PD-L1 expression on both tumor cells and tumor-associated stroma cells. PD-L1 was defined as "positive" if CPS was ≥ 1 and "negative" or not expressed if CPS <1.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!