Human Granzyme B Antibody

Detection of Human Granzyme B by Simple Western^TM.

Simple Western lane view shows lysates of NK-92 human natural killer lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Granzyme B at approximately 42 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Granzyme B Monoclonal Antibody (Catalog # MAB2906) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Granzyme B in human tonsil.

Granzyme B was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human Granzyme B Monoclonal Antibody (Catalog # MAB2906) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human Granzyme B by Simple Western

Patient derived NK cells can be expanded in vitro and NKEVs contain protein and nucleic acid cargo reflecting the cells of origin. NK cells were isolated from patient PBMCs via negative selection and EVs were isolated with size exclusion chromatography from the NK cell culture medium (n=20, 10 �pre� and 10 �post) (a). Representative image of cytokine profiling (n=12) of the secretome showed the presence of classic chemokines (CCL1, -5) and cytokines (IFNy, GM-CSF) (b). NKEVs were 100�200 nm in diameter post SEC (n=20) (c) and express canonical EV markers and cytotoxic NK proteins (d). Whole transcriptome sequencing (n=21; 20 patient NKEV and 1 control pool NKEV) found the majority of NKEV RNA cargo is protein coding and long noncoding transcripts (e). Differential gene expression (DE) analysis of LUAD/LUSC NKEVs identified a small number of significantly DE transcripts between LUAD and LUSC, such as ERAP2, which is upregulated in LUAD NKEVs (f). Mass spectrometry proteomics analysis (n=21; 20 patient NKEV and 1 control pool NKEV) of NKEV cargo identified over 4000 proteins (g), and modestly differentially expressed proteins between LUAD and LUSC groups (h). (b) is one representative dot blot, PT007-post and (d) is one representative Western blot, PT007-post. (c) shows NTA analysis for all individual samples in grey, with the mean distribution in red and SEM error bars. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/40821787/), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Granzyme B by Simple Western

Patient derived NK cells can be expanded in vitro and NKEVs contain protein and nucleic acid cargo reflecting the cells of origin. NK cells were isolated from patient PBMCs via negative selection and EVs were isolated with size exclusion chromatography from the NK cell culture medium (n=20, 10 “pre” and 10 “post) (a). Representative image of cytokine profiling (n=12) of the secretome showed the presence of classic chemokines (CCL1, -5) and cytokines (IFNy, GM-CSF) (b). NKEVs were 100–200 nm in diameter post SEC (n=20) (c) and express canonical EV markers and cytotoxic NK proteins (d). Whole transcriptome sequencing (n=21; 20 patient NKEV and 1 control pool NKEV) found the majority of NKEV RNA cargo is protein coding and long noncoding transcripts (e). Differential gene expression (DE) analysis of LUAD/LUSC NKEVs identified a small number of significantly DE transcripts between LUAD and LUSC, such as ERAP2, which is upregulated in LUAD NKEVs (f). Mass spectrometry proteomics analysis (n=21; 20 patient NKEV and 1 control pool NKEV) of NKEV cargo identified over 4000 proteins (g), and modestly differentially expressed proteins between LUAD and LUSC groups (h). (b) is one representative dot blot, PT007-post and (d) is one representative Western blot, PT007-post. (c) shows NTA analysis for all individual samples in grey, with the mean distribution in red and SEM error bars. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40821787), licensed under a CC-BY license. Not internally tested by R&D Systems.