Paraffin blocks were cut into 1 µm-thick sections and fixed onto a poly-L-lysine treated glass slide and then used for histological and immunohistochemical analyses. All sections were stained with H&E following manufacturer’s protocol to achieve correct orientation of the biopsy. Immunohistochemical staining was performed using Bond Polymer refine Detection kit on a BOND-MAX Automated IHC/ISH Stainer (Leica Biosystems) following the manufacturer’s protocol. The following antibodies were used: mouse monoclonal anti-human CD19 (NCL-L-CD19-163, Leica Biosystems), rat anti-human CD3 (MCA1477,Bio-Rad), mouse monoclonal anti-human CD4 (NCL-L-CD4-368, Leica Biosystems), mouse monoclonal anti-human CD8 (NCL-L-CD8-4B11, Leica Biosystems), mouse monoclonal anti-human Claudin-1 (ab56417, Abcam), mouse monoclonal anti-human E-Cadherin (36B5) (PA0387, Leica Biosystems), mouse monoclonal anti-human FoxP3 (ab22510, Abcam), mouse monoclonal anti-human γδ-TCR (sc-100289, Santa Cruz Biotechnology), mouse monoclonal anti-human Langerin (ab49730, Abcam), rabbit recombinant monoclonal [EPR20992] to Occludin (ab216327, Abcam), and rabbit polyclonal to Neutrophil Elastase (ab68672, Abcam). Positive and negative controls were included for each experiment.
Ncl l cd4 368
Manufactured by Leica
Sourced in United States
The NCL-L-CD4-368 is a laboratory equipment product. It is designed for use in scientific research and analysis. The core function of this product is to perform specific tasks related to the research and study of CD4 cells.
Automatically generated - may contain errors
Lab products found in correlation
A0452, by Agilent Technologies (2 mentions)
Ab22510, by Abcam (1 mentions)
Ab56417, by Abcam (1 mentions)
Ab68672, by Abcam (1 mentions)
Ab216327, by Abcam (1 mentions)
Bond max automated ihc ish stainer, by Leica (1 mentions)
Ncl l cd8 4b11, by Leica (1 mentions)
Sc 100289, by Santa Cruz Biotechnology (1 mentions)
Mca1477, by Bio-Rad (1 mentions)
Bond polymer refine detection kit, by Leica (1 mentions)
N1580, by Agilent Technologies (1 mentions)
M0876, by Agilent Technologies (1 mentions)
Dako pre treatment module, by Agilent Technologies (1 mentions)
Dako envision flex, by Agilent Technologies (1 mentions)
Dako envision flex kit, by Agilent Technologies (1 mentions)
Jcb117, by Agilent Technologies (1 mentions)
Bond max, by Leica (1 mentions)
Mcl 1, by Santa Cruz Biotechnology (1 mentions)
Pd l1 clone e1l3n, by Cell Signaling Technology (1 mentions)
Gapdh mab374, by Merck Group (1 mentions)
9 protocols using ncl l cd4 368
1
Comprehensive Immunohistochemical Profiling of Tissues
2
Immunohistochemical Analysis of Endomyocardial Biopsies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Buffered formalin-fixed and paraffin-embedded (FFPE) tissues obtained from endomyocardial biopsies were subjected to immunohistochemistry staining. FFPE tissues were cut into 4-μm-thick sections and stained with hematoxylin, eosin, and Masson’s trichrome stains for morphological observation. The immunohistochemical study was performed using primary antibodies against CD3 (#N1580, Dako, Glostrup, Denmark) for T lymphocytes, CD4 (NCL-L-CD4-368, Leica Biosystems, Wetzlar, Germany) for helper T lymphocytes, CD8 (NCL-CD8-4B11, Leica Biosystems) for cytotoxic T lymphocytes, and CD68 (M0876, Dako) for macrophages. In addition, a commercially available tenascin-C antibody (4F10TT, #10337, Immuno-Biological Laboratory Co., Ltd., Gunma, Japan) was also used. We used BOND-III automated immunohistochemical staining system (Leica Microsystems K.K., Tokyo, Japan). After immunostaining, CD3-, CD8-, and CD68-positive cells were counted by two pathologists in a high-power field in the most intensively lymphocyte-infiltrated area. Each sample was classified using the grading system reported by Palaskas et al. [13 (link)].
3
Immunohistochemical Analysis of Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FFPE pUM and mUM samples were sectioned at 4 μm thickness and underwent antigen retrieval using the Dako pretreatment module (Agilent Technologies UK Ltd, Stockport, UK); slides were then incubated in a high‐pH bath containing Tris/EDTA buffer, pH 9.0 (Dako EnVision™ FLEX, Agilent) at 96°C for 20 min. IHC was performed using a Dako Autostainer PLUS machine, using the Dako EnVision™ FLEX Kit (Agilent) according to the manufacturer's instructions. Slides were incubated with the following antibodies for 30 min: BAP1 (cat. No sc‐28 383/C‐4, dilution 1:200; Santa Cruz Biotechnology, Dallas, TX, USA), CD3 (cat. No IR503/polyclonal, ready to use; Dako Cytomation, CA, USA), CD4 (cat. No NCL‐L‐CD4/368, dilution 1:20; Leica Biosystems, Lincolnshire, IL, USA), CD8 (cat. No M7103/ C8/144B, dilution 1:200; Dako), CD163 (NCL‐L‐CD163/10D6, dilution 1:400; Leica Biosystems), and CD38 (NCL‐L‐CD38‐290/SPC32, dilution 1:100; Leica Biosystems).
The sections were counterstained with haematoxylin. Additional sections were treated with isotype controls at the same concentration as the primary antibodies.
The sections were counterstained with haematoxylin. Additional sections were treated with isotype controls at the same concentration as the primary antibodies.
4
Immunohistochemical Analysis of Lymphoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Biopsies were fixed in 10% neutral buffered formalin, embedded in paraffin, and stained with hematoxylin-eosin. Immunohistochemical staining and in situ hybridization were performed on an automated stainer (BOND-MAX; Leica Biosystems). The presence of EBV was demonstrated by in situ hybridization with a probe for small RNA–encoding regions 1 and 2 (EBERs) probe (Dako). Antibodies used were anti-CD3 (polyclonal rabbit, CD3ε, A0452; Dako, Agilent Technologies), CD4 (monoclonal IgG1k mouse, NCL-L-CD4-368; Leica Biosystems), CD8 (monoclonal mouse IgG1k, C8/144B; Dako), CD20 (monoclonal mouse IgG2ak, L26; Dako), and anti-CD79a (monoclonal mouse IgG1k, JCB117; Dako).
5
Immunoblot and Immunohistochemistry Analyses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies and concentrations used: pThr18/Ser19-MLC2 (#3674; 1:750, immunoblot), pSer19-MLC2 (#3671; 1:50, immunohistochemistry; 1:200, immunofluorescence), MLC2 (#3672; 1:750), pT202/Y204-p44/42 (ERK1/2) (#4370; 1:1,000), pY705-STAT3 (#9145; 1:750), PD-L1 (clone E1L3N, #13684, 1:200) from Cell Signaling Technology; STAT3 (sc-482; 1:500), ERK2 (sc-154; 1:1,000), MCL-1 (sc-819; 1:1,000), GFP (sc-8334; 1:1,000) from Santa Cruz Biotechnology; GAPDH (MAB374; 1:10,000) from Millipore; P-H2A.X (S139) (ab2893;1:1000), CD206 (ab64693; 1:1,000), CD3 (anti-mouse, ab134096; 1:500), CD4 (anti-mouse, clone I3T4, ab183685; 1:300), FoxP3 (anti-human, clone 236A/E7, ab20034; 1:200) from Abcam; F4/80 (anti-mouse, clone BM8, MF48000, 1:1000), CD8a (anti-mouse, clone Ly2, 14-0808-82; 1:200), FoxP3 (anti-mouse, clone FJK-16s, 14-5773-82; 1:200) from Invitrogen; CD4 (anti-human, clone 11E9, NCL-L-CD4-368; 1:300) from Novocastra.
6
Immunohistochemical Profiling of T-Cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antigen retrieval was performed on freshly cut sections using a decloaking chamber for 5 min at 125 °C in TRIS buffer (pH 9.0). Endogenous peroxidase was blocked by incubation with 3% peroxide at room temperature for 8 min. IHC was performed using the following antibodies:
• Pan T-cells were detected with a CD3 antibody (Dako A0452) at a 1:200 dilution. • T-helper cells were detected with a CD4 antibody (Novocastra NCL-L-CD4-368) at a 1:100 dilution.
• Cytotoxic T cells were detected with a CD8 antibody (Dako M7103) at a 1:100 dilution. • T regulatory cells (Tregs) were detected with a FOXP3 antibody (Thermo Fisher Scientific 14-4777-82) at a 1:100 dilution.
• Pan T-cells were detected with a CD3 antibody (Dako A0452) at a 1:200 dilution. • T-helper cells were detected with a CD4 antibody (Novocastra NCL-L-CD4-368) at a 1:100 dilution.
• Cytotoxic T cells were detected with a CD8 antibody (Dako M7103) at a 1:100 dilution. • T regulatory cells (Tregs) were detected with a FOXP3 antibody (Thermo Fisher Scientific 14-4777-82) at a 1:100 dilution.
7
Immunohistochemical Analysis of Immune Markers in Glioblastoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry was performed on formalin fixed paraffin-embedded (FFPE) tissue using the avidin-biotin-peroxidase complex method according to the manufacturer's protocol (Vectastain, PK-4004, Vector Laboratories; Burlingame, CA, USA). FFPE sections from GBM patients (n = 158) were immunostained with the following antibodies according to standard protocols: mouse anti-human cytomegalovirus clone CCH2+ DDG9 detecting IE-1 protein (IR75261-2), rabbit anti-human CD3 (A0452, Dako; Carpinteria, CA, USA), CD4 (NCL−L-CD4-368, Novocastra), CD8α (M7103, Dako), and mouse anti-human NKp46 (LS-B2105/10193, BioSite; San Diego, CA). Human tonsil tissue was used for a positive control, and primary antibody (MOC-31, sc-52344, Santa Cruz Biotechnology; Dallas, TX, USA) was used as the negative control. For IE-1, placental tissue from a known HCMV+ patient was used as the positive control. CD3+,CD4+ and CD8+ cells were quantified by morphometry, using NIS-Elements BR v4 software (400 × magnification, Nikon), and the results were presented as a percentage of the total number of positive cells in a minimum of 4 randomly selected fields representing hot spots for each section analyzed. Tumors were designated positive or negative for HCMV antigens based on the presence or absence of immunolabelled cells.
8
Balloon-Assisted Endoscopy for Intestinal Abnormalities
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For patients with persisting symptoms in which the CE examination identified severe significant abnormalities, a subsequent balloon-assisted endoscopy (BAE) was performed as a therapeutic intervention or for diagnostic biopsy, where necessary. To reveal the association between villous atrophy and HIV infection, the expression of intestinal CD4+ T cells was examined using duodenal specimens and immunohistochemistry staining (NCL-L-CD4-368; Novocastra, Newcastle, United Kingdom).
9
Immunohistochemical Analysis of Tumor Immune Infiltrates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FUS-targeted peritumoral or tumoral tissues were collected during tumor resection 7 days after sonication. Paraformaldehyde-fixed and paraffin-embedded methods were used to prepare 4-μm-thick sections for IHC analysis. Anti-CD4 antibody (Novocastra, NCL-L-CD4-368) was used to identify helper T lymphocytes (CD3+/CD4+ HTL); anti-CD8 antibody (Abcam, ab17147) was used to specifically bind to cytotoxic T lymphocytes (CD3+/CD8+ CTL). FOXP3 marker (eBiocsience, 14-4777) was used for regulatory T cells (Tregs). Anti-CD68 antibody (Abcam, ab955) was applied for macrophages.
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!