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 cd8 4b11
Manufactured by Leica
Sourced in United Kingdom
The NCL-L-CD8-4B11 is a laboratory equipment product designed for research purposes. It functions as an antibody that specifically binds to the CD8 protein, which is expressed on the surface of certain T cells. This product can be used in various immunological and cell biology applications that require the identification or isolation of CD8-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
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 cd4 368, 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)
Envision hrp kit, by Agilent Technologies (1 mentions)
Hpa077312, by Merck Group (1 mentions)
Cd8 clone 4b11, by Leica (1 mentions)
Cd68 clone kp1, by Agilent Technologies (1 mentions)
Pd l1 clone 22c3, by Agilent Technologies (1 mentions)
Granzyme b, by Akoya Biosciences (1 mentions)
3 protocols using ncl l cd8 4b11
1
Comprehensive Immunohistochemical Profiling of Tissues
2
Quantification of Exosomal DOK3, CD19, and CD8 in Paraffin Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin‐embedded specimens were prepared from patients who showed a high or low level of plasma exosomal DOK3 (Exo‐DOK3). The slides were treated with xylene and ethanol to remove the paraffin. The ENVISION+ Kit/HRP (DakoCytomation,) was used to detect DOK3, CD19, and CD8. After blocking the endogenous peroxidase activity and nonspecific protein staining, affinity‐purified rabbit anti‐DOK3 polyclonal antibody (HPA077312; Sigma‐Aldrich), mouse anti‐CD19 monoclonal antibody (BT51E; Leica Biosystems), or mouse anti‐CD8 monoclonal antibody (NCL‐L‐CD8‐4B11; Leica Biosystems) were added as the primary antibody. Then, slides were treated with horseradish peroxidase (HRP)‐labeled anti‐rabbit IgG for DOK3 staining or anti‐mouse IgG for CD19 and CD8 staining, respectively. Finally, chromogen 3,3′‐diaminobenzidine (DAB) was added as the substrate. Serial sections of the tissue specimens were counterstained with H&E. Quantification of cells with positive staining was performed using ImageJ software. The “Brightness” of “Threshold Color” was set from 111 to 170 to defined DAB‐stained cells for all slides. Then the area of the defined DAB‐stained cells was measured within ~1.7 mm2 of tissue sections. Finally, the positive area per unit tissue section (μm2/mm2) was calculated and shown.
3
Multiplex IHC Profiling of Tumor-Infiltrating Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical (IHC) staining using whole section FFPE tissues was performed with the following primary antibodies: CD8 (clone 4B11, 1:400, Cat#NCL-L-CD8-4B11, Leica Biosystems, Newcastle, UK), granzyme B (clone 11F1, 1:128, Cat#NCL-L-GRAN-B, Leica Biosystems, Newcastle, UK), CD68 (clone KP1, 1:2000, Cat#M0814, Dako, Glostrup, Denmark), and PD-L1 (clone 22C3, 1:50, Cat#M3653, Dako, Glostrup, Denmark). PD-L1 expression was evaluated as the tumor proportion score (TPS), which was defined as the percentage of tumor cells with PD-L1 expression, and the combined positive score (CPS), which was defined as the number of PD-L1-expressing tumors and immune cells (lymphocytes and macrophages) divided by the number of all tumor cells and multiplied by 100. To quantify CD8-, CD68-, and granzyme B-positive immune cells, up to 10 representative images of each IHC slide were acquired and analyzed by two expert genitourinary pathologists (S.H. and G.Y.K.) using the inForm 2.6 software (Akoya Biosciences, Marlborough, MA, USA). Positive cells were scored out of the total number of cells counted in representative images.
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!