polyclonal antibody against cd68  (Millipore)

Journal: Scientific Reports

Article Title: KPC-luciferase-expressing cells elicit an anti-tumor immune response in a mouse model of pancreatic cancer

doi: 10.1038/s41598-024-64053-0

Figure Lengend Snippet: KPC-Luc tumors have increased immune cell infiltration when compared to KPC tumors. Immunofluorescence staining of immune cell infiltration in the tumor (red). Nuclei are stained by DAPI (blue). White arrows point to positive stainings. ( A ) Representative images of immunofluorescence staining of KPC and KPC-Luc tumors 9 and 13 days after cell implantation, showing infiltration of CD8 + T cells. ( B ) Quantitative analysis depicted a significantly increased number of CD8 + T cells infiltrated in the tumors of KPC-Luc cells implanted mice, when compared to KPC cells implanted mice, at 13 days of tumor growth, but not at 9 days. Moreover, in the KPC-Luc group, tumor-infiltrated CD8 + T cells increased between days 9 and 13 after cell implantation; n = 3. ( C ) Representative images of immunofluorescence staining of KPC and KPC-Luc tumors 9 and 13 days after cell implantation, showing infiltration of CD68 + cells (macrophages). ( D ) Quantitative analysis showed no significant differences in the number of CD68 + cells infiltrated in the tumor of KPC and KPC-Luc implanted mice at 9 or 13 days of tumor growth; n = 3; Data is presented as mean ± SD. Two-way ANOVA followed by Sidak’s multiple comparisons; *p < 0.01. Scale bars in ( A ) and ( C ) represent 50 µm.

Article Snippet: For this purpose, slides were deparaffinized with xylol and rehydrated with increasing concentrations of EtOH, blocked with fish serum (37527, Invitrogen) for 20 min and incubated overnight with rabbit primary monoclonal antibody against CD8b (1:500 dilution, ab228965, Sigma) or polyclonal antibody against CD68 (1:500 dilution, ab125212, Sigma), followed by 1 h incubation with a goat anti-rabbit secondary antibody conjugated with Alexa Fluor 647 (1:200 dilution, a21244, Invitrogen), and counterstained with DAPI for 30 min for nuclei staining.

Techniques: Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: KPC-luciferase-expressing cells elicit an anti-tumor immune response in a mouse model of pancreatic cancer

doi: 10.1038/s41598-024-64053-0

Figure Lengend Snippet: After regression, KPC-Luc tumors do not regrow. ( A ) Mice were injected orthotopically with 5 × 10 4 KPC-Luc cells and monitored for tumor growth once or twice per week by IVIS Spectrum. 70 days after cell implantation, mice were sacrificed and analyzed for the presence of tumors. ( B ) Representative images of bioluminescent tumors are shown 7 and 70 days after tumor cell implantation. The majority of mice lost the bioluminescence signal by day 70. ( C ) Graph showing bioluminescence signals of 5 individual mice over time. The signal was present until the second-week post-implantation before it decreased. In four out of five mice, no bioluminescence signal could be detected from day 3 to 70 after cell implantation. Only one mouse had a constant increase of signal from 20 days after tumor cell injection that persisted until the day of sacrifice. ( D ) Immunofluorescence staining showed infiltration of CD8 + T cells and CD68 + cells in a representative section of a tumor obtained 70 days after KPC-Luc implantation. White arrows point to positive staining of the immune cells. Scale bars in ( D ) represent 50 µm.

Article Snippet: For this purpose, slides were deparaffinized with xylol and rehydrated with increasing concentrations of EtOH, blocked with fish serum (37527, Invitrogen) for 20 min and incubated overnight with rabbit primary monoclonal antibody against CD8b (1:500 dilution, ab228965, Sigma) or polyclonal antibody against CD68 (1:500 dilution, ab125212, Sigma), followed by 1 h incubation with a goat anti-rabbit secondary antibody conjugated with Alexa Fluor 647 (1:200 dilution, a21244, Invitrogen), and counterstained with DAPI for 30 min for nuclei staining.

Techniques: Injection, Immunofluorescence, Staining

Journal: BMC Genomics

Article Title: Differential gene expression and immune cell infiltration in maedi-visna virus-infected lung tissues

doi: 10.1186/s12864-024-10448-2

Figure Lengend Snippet: Immunostaining of CD68-positive macrophages and CD19‐positive B cells infiltrating the lung. ( A ) Control samples for CD68 immunostaining. ( B ) CD68 immunostaining of lungs from Maedi-visna virus (MVV)-infected sheep. ( C ) Control samples for CD19 immunostaining. ( D ) CD19 immunostaining of lungs from MVV-infected sheep (red bar, 50 μm; green bar, 40 μm)

Article Snippet: The tissues were incubated with a 1:200 dilution of CD4 polyclonal antibody (19068-1-AP, Proteintech, Wuhan, China), rabbit polyclonal CD8 alpha (ab4055, Abcam, Cambridge, USA), CD19 rabbit mAb (A19013, ABclonal Technology Co., Ltd.), and rabbit polyclonal antibody against CD68 (DF7518, Affinity Biosciences) overnight at 4 °C.

Techniques: Immunostaining, Control, Virus, Infection

Journal: Journal of Translational Medicine

Article Title: Parathyroid hormone stimulates bone regeneration in an atrophic non-union model in aged mice

doi: 10.1186/s12967-023-04661-y

Figure Lengend Snippet: A Representative immunohistochemical images of CD31-positive microvessels (arrowheads) within the callus tissue of controls and PTH-treated mice at 2 and 10 weeks after surgery. Scale bars: 25 μm. B CD31-positive microvessels/HPF within the callus tissue of controls (white bars, n = 5 at 2 weeks after surgery, n = 9 at 10 weeks after surgery) and PTH-treated mice (black bars, n = 5 at 2 weeks after surgery, n = 9 at 10 weeks after surgery) at 2 and 10 weeks after surgery, as assessed by immunohistochemical analysis. Mean ± SEM; *p < 0.05 vs. control. C Representative immunohistochemical images of MPO-positive cells (arrowheads) within the callus tissue of controls and PTH-treated mice at 2 and 10 weeks after surgery. Scale bars: 25 μm. D MPO-positive cells/HPF within the callus tissue of controls (white bars, n = 5 at 2 weeks after surgery, n = 9 at 10 weeks after surgery) and PTH-treated mice (black bars, n = 5 at 2 weeks after surgery, n = 9 at 10 weeks after surgery) at 2 and 10 weeks after surgery, as assessed by immunohistochemical analysis. Mean ± SEM; *p < 0.05 vs. control. E Representative immunohistochemical images of CD68-positive cells (arrowheads) within the callus tissue of controls and PTH-treated mice at 2 and 10 weeks after surgery. Scale bars: 25 μm. F CD68-positive cells/HPF within the callus tissue of controls (white bars, n = 5 at 2 weeks after surgery, n = 9 at 10 weeks after surgery) and PTH-treated mice (black bars, n = 5 at 2 weeks after surgery, n = 9 at 10 weeks after surgery) at 2 and 10 weeks after surgery, as assessed by immunohistochemical analysis. Mean ± SEM; *p < 0.05 vs. control

Article Snippet: To detect the neutrophilic granulocyte marker myeloperoxidase (MPO) and the macrophage marker CD68, sections were stained with a polyclonal rabbit anti-mouse antibody against MPO (1:100; Abcam) and a polyclonal rabbit anti-mouse antibody against CD68 (1:100; Abcam).

Techniques: Immunohistochemistry