rabbit anti human her2 monoclonal antibody  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Flow cytometry for HER2-positive and EGFR-positive cells in populations of SK-OV-3 and/or MDA-MB-468 cells SK-OV-3 or MDA-MB-468 cells or mixtures of these cells immunostained with (A) anti-HER2 Alexa Fluor 488-conjugated trastuzumab or (B) with anti-EGFR Alexa Fluor 647-conjugated panitumumab were analyzed by flow cytometry. The proportion of HER2-positive or EGFR-positive cells was quantified as the area under the relevant peak for SK-OV-3 or MDA-MB-468 cells, respectively and plotted vs. the percentage of (C) SK-OV-3 cells or (D) MDA-MB-468 cells in the cell population.

Article Snippet: Rabbit anti-human HER2 monoclonal antibody , ThermoFisher/Invitrogen , #SP3.

Techniques: Flow Cytometry

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Flow cytometry for HER2-positive cells in dissociated tumors composed of SK-OV-3 and/or MDA-MB-468 cells Tumor xenografts in NOD/SCID mice were dissociated at 6 weeks post s.c. inoculation of tumor cells then the recovered cells were immunostained with anti-HER2 Alexa Fluor 488-conjugated trastuzumab and analyzed by flow cytometry. (A) Based on gating thresholds, MDA-MB-468 tumors were assigned as 4.2% HER2-positive cells and 95.8% HER2-negative cells. (B) SK-OV-3 tumors were assigned as 84.6% HER2-positive cells and 15.4% HER2-negative cells. (C) Replicate tumors ( n = 3) formed by the inoculation of 30% SK-OV-3 cells and 70% MDA-MB-468 cells were assigned as 40.8%, 23.1%, and 28.1% HER2-positive cells and 59.2%, 76.9%, and 71.9% HER2-negative cells.

Article Snippet: Rabbit anti-human HER2 monoclonal antibody , ThermoFisher/Invitrogen , #SP3.

Techniques: Flow Cytometry

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Immunohistochemical (IHC) staining of tumors composed of SK-OV-3 and/or MDA-MB-468 cells for HER2-positive and EGFR-positive cells Representative sections of tumors formed by the inoculation of (A) SK-OV-3 cells or (B) MDA-MB-468 cells or (C) a mixture of 30% SK-OV-3 cells and 70% MDA-MB-468 cells at 6 weeks post-inoculation in NOD/SCID mice. Sections were stained with hematoxylin and eosin (H&E) or immunostained for HER2 or EGFR. Sections are shown at 4× or 20× amplification. (D) The percentage of cells immunopositive for HER2 or EGFR in replicate tumor sections was quantified by image analysis and is shown for SK-OV-3 or MDA-MB-468 tumors or tumors formed by inoculating a mixture of 30% SK-OV-3 cells and 70% MDA-MB-468 cells. Data shown are mean ± SD ( n = 3). Statistically significant differences (Student’s t test; p < 0.05) are indicated by the asterisks.

Article Snippet: Rabbit anti-human HER2 monoclonal antibody , ThermoFisher/Invitrogen , #SP3.

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, Amplification

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: PET/CT images of NOD-SCID mice with s.c. SK-OV-3 tumors with homogeneous HER2 expression or tumors with heterogeneous HER2 expression formed from a mixture of SK-OV-3 and MDA-MB-468 cells Representative images (supine position) of NOD-SCID mice with s.c. SK-OV-3 tumors on the right flank that homogeneously express HER2 (blue arrow) and tumors formed by the inoculation of a mixture of SK-OV-3 cells (30%) and MDA-MB-468 cells (70%) with heterogeneous HER2 and EGFR expression on the left flank (red arrow). Images were obtained at 24 h and 48 h p.i. of (A) [ 64 Cu]Cu-NOTA-trastuzumab Fab-PEG 24 -EGF, (B) [ 64 Cu]Cu-NOTA-trastuzumab Fab or (C) [ 64 Cu]Cu-NOTA-EGF. All images were adjusted to the same intensity and the intensity scale (ranging from 1.5% ID/g to 12.5% ID/g) is shown at the right of each set of images. Normal organ uptake is visible on the 24 h and 48 h p.i. images but is labeled on the 48 h p.i. images. L: liver; K: kidneys; Sp: spleen.

Article Snippet: Rabbit anti-human HER2 monoclonal antibody , ThermoFisher/Invitrogen , #SP3.

Techniques: Positron Emission Tomography-Computed Tomography, Expressing, Labeling

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: PET/CT images of NOD-SCID mice with s.c. MDA-MB-468 tumors with homogeneous EGFR expression or tumors with heterogeneous EGFR expression formed from a mixture of MDA-MB-468 and SK-OV-3 cells Representative images (supine position) of NOD-SCID mice with s.c. MDA-MB-468 tumors on the right flank that homogeneously express EGFR (blue arrow) and tumors formed by the inoculation of a mixture of MDA-MB-468 cells (70%) and SK-OV-3 cells (30%) with heterogeneous HER2 and EGFR expression on the left flank (red arrow). Images were obtained at 24 h and 48 h p.i. of (A) [ 64 Cu]Cu-NOTA-trastuzumab Fab-PEG 24 -EGF, (B) [ 64 Cu]Cu-NOTA-trastuzumab Fab or (C) [ 64 Cu]Cu-NOTA-EGF. All images were adjusted to the same intensity and the intensity scale (ranging from 1.5% ID/g to 12.5% ID/g) is shown at the right of each set of images. Normal organ uptake is visible on the 24 h and 48 h p.i. images but is labeled on the 48 h p.i. images. L: liver; K: kidneys; Sp: spleen.

Article Snippet: Rabbit anti-human HER2 monoclonal antibody , ThermoFisher/Invitrogen , #SP3.

Techniques: Positron Emission Tomography-Computed Tomography, Expressing, Labeling

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet:

Article Snippet: Rabbit anti-human HER2 monoclonal antibody , ThermoFisher/Invitrogen , #SP3.

Techniques: Recombinant, Expressing, Software, Positron Emission Tomography, Computed Tomography, Flow Cytometry

Journal: Oncology Research

Article Title: Investigation on the Anti-Cancer Effects of HER2-Targeted CAR-T Cells Engineered Using the PiggyBac Transposon System

doi: 10.32604/or.2025.065394

Figure Lengend Snippet: The optimization of the ratio of transposon plasmid and transposase plasmid for transfection of anti-HER2 and anti-HER2-13 CAR-T cells. ( A ) The schematic representation of the anti-HER2/HER2-13 CAR structure. ( B ) Diagram of the construction and expansion of CAR-T cells. On day 0, the isolated PBMC were electroporated with PiggyBac transposase plasmid along with transposon DNA plasmids encoding the PB-HER2(13)-CAR-puro. On day 1, magnetic beads and IL-2, IL-7 and IL-21 were added for expansion for 3–5 days, then the cells were followed by removal of the magnetic beads and continued culture for approximately 18–21 days. ( C , D ) The representative flow cytometry graphs of CAR + expression in CAR-T conducted with three different ratios (μg:μg) of transposon plasmid and transposase plasmid. ( E ) The analytic results of the total cell proliferation, CD3 + CAR + , CD4 + CAR + , CD8 + CAR + , CD4 + CAR + /CD8 + CAR + and CD3 + CAR + cell numbers. The data was from two different donors

Article Snippet: They were then incubated overnight at 4°C with monoclonal rabbit anti-human HER2 antibody (Cell Signaling Technology, 4290S, Danvers, MA, USA, 1:1000) or anti-β-actin mAb (mouse origin) (Proteintech, 66009-1, Rosemont, IL, USA, 1:1000).

Techniques: Plasmid Preparation, Transfection, Isolation, Magnetic Beads, Flow Cytometry, Expressing

Journal: Oncology Research

Article Title: Investigation on the Anti-Cancer Effects of HER2-Targeted CAR-T Cells Engineered Using the PiggyBac Transposon System

doi: 10.32604/or.2025.065394

Figure Lengend Snippet: The differences in in vitro expansion abilities, CD3 + CAR + , CD4 + CAR + /CD8 + CAR + expression between anti-HER2 and anti-HER2-13 CAR-T cells. ( A ) The proliferation of CAR-T cells or NT cells which were transfected with empty vector (PB-puro). The arrow indicated the different time points for adding puromycin. The data was from three different donors. ( B ) On the 7th and 14th day, respectively, the representative images of CAR-T cells constructed with two sequences of scFv were taken at a magnification of 200× to compare their morphology. The scale bars in the pictures are 50 μm. ( C ) Representative flow cytometry photos of CD3 + CAR + , CD4 + CAR + and CD8 + CAR + expression in the CAR-T cells and NT cells. ( D – G ) The analytic results of CD3 + CAR + , CD4 + CAR + , CD8 + CAR + and CD4 + CAR + /CD8 + CAR + cells. The data was from five (or seven) different donors. The difference between the anti-HER2 group and anti-HER2-13 group was determined by Student t -test, * p < 0.05; ns (not significant, p ≥ 0.05), n = 5

Article Snippet: They were then incubated overnight at 4°C with monoclonal rabbit anti-human HER2 antibody (Cell Signaling Technology, 4290S, Danvers, MA, USA, 1:1000) or anti-β-actin mAb (mouse origin) (Proteintech, 66009-1, Rosemont, IL, USA, 1:1000).

Techniques: In Vitro, Expressing, Transfection, Plasmid Preparation, Construct, Flow Cytometry

Journal: Oncology Research

Article Title: Investigation on the Anti-Cancer Effects of HER2-Targeted CAR-T Cells Engineered Using the PiggyBac Transposon System

doi: 10.32604/or.2025.065394

Figure Lengend Snippet: The differences in memory (CD44, CD62L) and characteristic markers of T cell exhaustion (PD-1, LAG-3, TIM-3) between anti-HER2 and anti-HER2-13 CAR-T cells. ( A ) Representative flow cytometry photos of memory and exhaustion marker expression in anti-HER2 and anti-HER2-13 CAR-T cells. ( B ) The analytic results of memory and exhaustion marker expression in CAR-T cell (0: fresh state). The data was from three different donors. ( C , D ) The influence on the cell viability, memory (CD44, CD62L) and key exhaustion indicators (LAG-3, TIM-3, PD-1) of anti-HER2 and anti-HER2-13 CAR-T cells under cryopreservation (0: fresh state, 1: one month, 6: six months). The data was from two different donors. The disparity between the anti-HER2 group and the anti-HER2-13 group was assessed through a Student’s t -test, which yielded ns (showing non-significant differentials, p ≥ 0.05)

Article Snippet: They were then incubated overnight at 4°C with monoclonal rabbit anti-human HER2 antibody (Cell Signaling Technology, 4290S, Danvers, MA, USA, 1:1000) or anti-β-actin mAb (mouse origin) (Proteintech, 66009-1, Rosemont, IL, USA, 1:1000).

Techniques: Flow Cytometry, Marker, Expressing

Journal: Oncology Research

Article Title: Investigation on the Anti-Cancer Effects of HER2-Targeted CAR-T Cells Engineered Using the PiggyBac Transposon System

doi: 10.32604/or.2025.065394

Figure Lengend Snippet: The anti-tumor effects of CAR-T cells on different HER2-positive or HER2-negative cancer cell lines. ( A ) The protein expression levels of HER2 in different tumor cells of lung adenocarcinoma (SPC-A-1 HER2+ /SPC-A-1) and breast cancer (MDA-MB-231 HER2+/ MDA-MB-231 and SKBR-3/SKBR-3 shHER2) were determined by western blotting, and the expression of β-actin was measured to confirm equal protein levels. ( B – D ) Cytotoxic effects of CAR-T cells on the above cancer cells. Cell viability was measured by colorimetric CCK-8 assay after treatment with CAR-T cells or NT cells for 24 h. The differences among CAR-T groups and NT on SPC-A-1 HER2+ /MDA-MB-231 HER2+ /SKBR-3 cells were determined by two-way ANOVA and post hoc Tukey multiple comparison tests. & p < 0.05, && p < 0.01, &&& p < 0.001 anti-HER2-13 CAR-T group vs. NT group; $ p < 0.05, $$$ p < 0.001 anti-HER2 CAR-T group vs. NT group. The CAR-T groups on SPC-A-1 HER2+ /SPC-A-1, MDA-MB-231 HER2+ /MDA-MB-231 and SKBR-3/SKBR-3 shHER2 cells were determined by two-way ANOVA and post hoc Tukey multiple comparison tests. # p < 0.05 anti-HER2-13 CAR-T+SPC-A-1 HER2 vs. anti-HER2-13 CAR-T+SPC-A-1, and anti-HER2-13 CAR-T+MDA-MB-231 HER2 vs. anti-HER2-13 CAR-T+MDA-MB-231; #### p < 0.0001 indicates the statistical differences between anti-HER2-13 CAR-T+SKBR-3 and anti-HER2-13 CAR-T+SKBR-3 shHER2 group; **** p < 0.0001 indicates the statistical differences between anti-HER2 CAR-T+SKBR-3 and anti-HER2 CAR-T+SKBR-3 shHER2 group. The line plots display the mean value ± standard deviation (SD) from biological replicates (n = 3 wells per experiment)

Article Snippet: They were then incubated overnight at 4°C with monoclonal rabbit anti-human HER2 antibody (Cell Signaling Technology, 4290S, Danvers, MA, USA, 1:1000) or anti-β-actin mAb (mouse origin) (Proteintech, 66009-1, Rosemont, IL, USA, 1:1000).

Techniques: Expressing, Western Blot, CCK-8 Assay, Comparison, Standard Deviation

Journal: Oncology Research

Article Title: Investigation on the Anti-Cancer Effects of HER2-Targeted CAR-T Cells Engineered Using the PiggyBac Transposon System

doi: 10.32604/or.2025.065394

Figure Lengend Snippet: The potential mechanisms involved in cytotoxicities of CAR-T cells against HER2 positive or negative lung adenocarcinoma and breast cancer cell lines. ( A – H ) Cytokines (IFN-γ, TNF-α, IL-10, IL-6, IL-2, GM-CSF) and effector molecule (perforin, granzyme B) production were quantified by ELISA following CAR-T cell co-culture with tumor targets at a 2:1 effector-to-target ratio. Compared with the NT control group, the statistical significance was assessed using two-way ANOVA followed by Dunnett’s post hoc test: * p < 0.05, ** p < 0.01, *** p < 0.001; the data were from two independent experiments with three wells each

Article Snippet: They were then incubated overnight at 4°C with monoclonal rabbit anti-human HER2 antibody (Cell Signaling Technology, 4290S, Danvers, MA, USA, 1:1000) or anti-β-actin mAb (mouse origin) (Proteintech, 66009-1, Rosemont, IL, USA, 1:1000).

Techniques: Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Control

Journal: Oncology Research

Article Title: Investigation on the Anti-Cancer Effects of HER2-Targeted CAR-T Cells Engineered Using the PiggyBac Transposon System

doi: 10.32604/or.2025.065394

Figure Lengend Snippet: The effects of anti-HER2, anti-HER2-13 CAR-T cells and trastuzumab on the MDA-MB-231 HER2+ xenograft mouse model. ( A ) The establishment and treatment pattern of the breast cancer MDA-MB-231 HER2+ mouse model. ( B ) The body weight of mice in each group was measured throughout the entire treatment period. ( C ) The representative photographic documentation of resected murine mammary tumors. ( D ) The tumor weight of dissected MDA-MB-231 HER2+ in mean ± SEM of 3–5 tumors. ### p < 0.001 anti-HER2 CAR-T+IL-2 group vs. anti-HER2-13 CAR-T+IL-2 group, *** p < 0.001, NT+IL-2 group vs. anti-HER2 CAR-T+IL-2 groups, by two-way ANOVA followed by post-hoc Tukey’s multiple comparison. * p < 0.05, NT+IL-2 group vs. anti-HER2-13 CAR-T+IL-2 group by Student t -test; n = 5 per group

Article Snippet: They were then incubated overnight at 4°C with monoclonal rabbit anti-human HER2 antibody (Cell Signaling Technology, 4290S, Danvers, MA, USA, 1:1000) or anti-β-actin mAb (mouse origin) (Proteintech, 66009-1, Rosemont, IL, USA, 1:1000).

Techniques: Comparison

Journal: Frontiers in Pharmacology

Article Title: Analysis of the heavy metal contents’ effect on steroidal saponins and the anti-breast cancer activity of Paris polyphylla var. yunnanensis

doi: 10.3389/fphar.2023.1277395

Figure Lengend Snippet: Effect of P. polyphylla var. yunnanensis from different origins on cancer cell proliferation and apoptosis induction in 4T1-tumor mice. (A) TUNEL staining (×400; n = 6). (B) Immunohistochemistry staining graph of Ki67 (×400; n = 6). (C) Immunohistochemistry staining graph of HER2 (×400; n = 6). (D) Immunohistochemistry staining graph of p53 (×400; n = 6). (E) Statistics of the TUNEL-positive cell. (F) Statistical analysis of Ki67 expression in 4T1-tumor mice. (G) Statistical analysis of p53 expression in 4T1-tumor mice. (H) Statistical analysis of p53 expression in 4T1-tumor mice. The results were expressed as mean ± SD, n = 6. ** p <0.01 and *** p <0.001.

Article Snippet: After 20 min of heat-induced antigen retrieval at 95°C, the serial sections were incubated overnight at 4°C with a combination of rabbit monoclonal anti-human p53 antibody (1:50 dilution; Abcam), rabbit monoclonal anti-human Ki67 antibody (1:500 dilution; Abcam), and rabbit monoclonal anti-human HER2 antibodies (1:500 dilution; Abcam).

Techniques: TUNEL Assay, Staining, Immunohistochemistry, Expressing