mouse gas6 duoset elisa kit  (R&D Systems)

Journal: Gynecologic Oncology Reports

Article Title: A fast-progressing orthotopic ovarian cancer model reveals synergistic antitumor effects of AXL-targeting nanobodies and Olaparib

doi: 10.1016/j.gore.2025.101998

Figure Lengend Snippet: AXL is highly expressed on ID8-derived cell lines. A. Stacked histogram plot showing expression of AXL on different passages of the ID8 cell lines compared to the FMO. B. Barplot showing the change in median fluorescence intensity of AXL (dMFI) in the different ID8 cell lines. Error bars indicate SEM. Statistical significance was determined using one-way ANOVA followed by post hoc Tukey’s multiple comparisons test. **, p < 0.01; ***, p < 0.001. C – H. Immunofluorescence images of the different ID8 cell lines at 40X magnification. Blue, DAPI; Red, AXL. I. Barplot showing the kinetic changes in the concentration of serum GAS6 (pg/mL) of P3 ID8 Thy1.1 tumor-bearing mice across weeks 1–6 post-inoculation and age-matched naïve mice. Error bars indicate SEM. Statistical significance was determined using one-way ANOVA followed by post hoc Tukey’s multiple comparisons test. *, p < 0.05.

Article Snippet: Quantification of the serum concentration of GAS6 was performed using the Mouse GAS6 DuoSet ELISA kit (R&D systems, cat# DY986) according to manufacturer instructions.

Techniques: Derivative Assay, Expressing, Fluorescence, Immunofluorescence, Concentration Assay

Journal: Gynecologic Oncology Reports

Article Title: A fast-progressing orthotopic ovarian cancer model reveals synergistic antitumor effects of AXL-targeting nanobodies and Olaparib

doi: 10.1016/j.gore.2025.101998

Figure Lengend Snippet: High expression of AXL and GAS6 mRNA in OC patients correlates with worse outcome. A-D. Kaplan-Meier plots showing the hazard ratio (HR) and logrank P value in OC patients for A. AXL mRNA for overall survival (OS) ( AXL- high: n = 169, AXL- low: n = 204); B. GAS6 mRNA for OS ( GAS6- high: n = 125, GAS6- low: n = 248); C. GAS6 mRNA for recurrence-free survival (RFS) ( GAS6- high: n = 122, GAS6- low: n = 55); D. mean expression of AXL and GAS6 mRNA for OS ( AXL and GAS6 -high: n = 203, AXL and GAS6- low: n = 170). E. Heatmap showing high AXL or GAS6 mRNA expression in different tissues from OC patients. F. Heatmap showing high AXL or GAS6 mRNA expression in platinum-resistant or sensitive OC patients.

Article Snippet: Quantification of the serum concentration of GAS6 was performed using the Mouse GAS6 DuoSet ELISA kit (R&D systems, cat# DY986) according to manufacturer instructions.

Techniques: Expressing

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: A) Mammary glands or tumors from wildtype, p53 −/− , or Balb/c mice containing PN1a or PN1b premalignant lesions were analyzed for Gas6 expression using ELISA. Recombinant Gas6 (rGas6) supplied by the manufacturer was used as positive control. Values shown (ng/ml of Gas6) are the mean and SEM from 3–6 independent experiments. ****p< 0.0001, **p< 0.01. B) Representative images of PN1a and PN1b tissues stained with a Gas6 antibody (n=3). Black arrowhead indicates Gas6 stromal staining and red arrowhead indicates Gas6 epithelial staining. Gas6 −/− mammary glands (MG) were used as a negative control. Scale bar= 20 μm. C) Cells were isolated from PN1a lesions at 8 weeks post-transplantation, FACS-sorted for CD45 + (CD45 + ), CD45 + CD3 + CD11b - (T cells), CD45 + F4/80 + CD11b + (macrophages) or CD45 - cells, and qPCR for Gas6 was performed. Graph depicts 2^ddCT after normalizing to 18S, and fold changes were calculated by setting the highest value (CD45 + ) equal to 1. Values shown are the mean and SD (n=3) from one representative experiment. D) LIN - epithelial cells were FACS-sorted from normal mammary glands (MEC) or PN1a and PN1b lesions (8 weeks post-transplantation) and relative expression of Axl, MerTK and Gas6 was detected by qPCR. Graph depicts 2^ddCT after normalizing to 18S values shown are the mean and SD (n=3) from one representative experiment. Fold changes were calculated after setting MEC equal to 1. ***p< 0.001 and ****p< 0.0001 E) Representative images of human DCIS stained with a Gas6 antibody displaying low and high epithelial Gas6 expression. Red arrowhead indicates Gas6 epithelial staining, while black arrowhead shows Gas6 stromal staining. Scale bar = 10 μm. F) Representative images of immunofluorescence staining showing Gas6 (red) and CD68 (green) co-localization in human DCIS. Scale bar = 20 μm.

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Recombinant, Positive Control, Staining, Negative Control, Isolation, Transplantation Assay, Immunofluorescence

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: Primary PN1a cells were cultured in Matrigel in the presence or absence of WT or Gas6 −/− BMDMs (PKH26-labelled). A) Representative phase contrast images (BMDMs: red) and confocal images of structures immunostained with antibodies to α6 integrin (red), CK18 (green), and CK14 (purple). Scale bar=50 μm. B) Quantification of the number of tumor-like (disorganized, grape-like morphology) and non-malignant colonies. At least 30 structures per group were examined for each experiment from 3 independent experiments. Error bars represent SEM, ****p<0.0001. C) ELISA for Gas6 levels in supernatants collected after 10 days of co-culture (13 days total culture). Error bars represent SEM from 3 independent experiments (**p=0.0012).

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

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

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: PN1a cells were cultured in Matrigel in the presence or absence of various concentrations of rGas6 for 13 days. A) Representative phase contrast and confocal images depicting morphological changes induced by Gas6 treatment. Cells were stained with antibodies against pAxl (green) or α6 integrin and nuclei were detected with DAPI (blue). Scale bar=50 μm. B) Quantification of non-malignant and tumor-like structures resulting from treatment with various concentrations of rGas6 or vehicle (PBS) control. The number of non-malignant or tumor-like structures from a minimum of 30 colonies per group were counted and expressed as a percent of total structures analyzed. Values shown are the mean and SEM from 3 independent experiments (p<0.0001). C) Graph depicts the presence or absence of pAxl staining within malignant structures after treatment with Gas6. Data are expressed as the percentage of total tumor-like structures (mean + SEM) from at least 30 colonies per group from 3 independent experiments (*p< 0.05).

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

Techniques: Cell Culture, Staining, Control

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: PN1a cells were cultured in Matrigel in the presence or absence of WT or Gas6 −/− BMDMs. Confocal images of structures stained with antibodies to A) pAxl (red), Axl (green) or B) pMerTK (red) are shown. Dapi staining was used to detect nuclei (blue). Arrow depicts pMerTK staining of a macrophage. Scale bar = 50 μm.

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

Techniques: Cell Culture, Staining

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: PN1a cells were cultured in Matrigel in the presence or absence of wildtype or Gas6 −/− BMDMs for a total of 13 days. A) Confocal images of cells stained with an antibody to cleaved CC3 and the percentage of CC3 + cells was quantified. B) Confocal images showing Ki-67 staining and graph depicts the percentage of proliferating cells in each group. Values from both graphs represent the mean and SEM per field of view from 3 independent experiments (*p< 0.05). Scale bar = 50 μm.

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

Techniques: Cell Culture, Staining

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: PN1a cells were cultured in Matrigel in the presence or absence of wildtype or Gas6 −/− BMDMs for 13 days, and then they were fixed and stained with antibodies to pAKT and pSTAT3. A) Representative confocal images of pAKT staining and quantification of pAKT + cells expressed as a percentage of total cells. B) Confocal images of pSTAT3 staining and quantification of pSTAT3 + cells expressed as a percentage of total cells. Nuclei were stained with DAPI (blue); scale bar = 50 μm. Values from both graphs represent the mean and SEM per field of view from 3 independent experiments (**p< 0.01 and *p< 0.05).

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

Techniques: Cell Culture, Staining

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: A) Representative images of 8 week PN1a (n=4), 18 week PN1a (n=3) and 12 week PN1b (n=3) tissues stained with an E-cadherin antibody. Scale bar = 20 μm. B) PN1a cells were cultured in Matrigel in the presence or absence of wildtype or Gas6 −/− BMDMs for 13 days, and then they were fixed and stained with an E-cadherin antibody (green) and nuclei were stained with DAPI (blue) (n=3). Representative confocal images of E-cadherin staining in PN1a cells are shown. Scale bar = 50 μm.

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

Techniques: Staining, Cell Culture

Journal: Oncogene

Article Title: Stromal Gas6 promotes the progression of premalignant mammary cells

doi: 10.1038/s41388-018-0593-5

Figure Lengend Snippet: PN1a cells were transplanted into the cleared fat pads of wildype or Gas6 −/− mice and allowed to progress to palpable tumors. Mice were euthanized when tumors reached 1 cm in diameter. A) Kaplan-Meier curve depicting the number of fat pads with palpable tumors at 0–35 weeks post-transplantation. 13 WT animals and 12 Gas6 −/− animals were analyzed. The log rank test was used for statistical analysis (p=0.01). Hazard ratio (Mantel Haenszel)=2.8 with 95% CI=1.2 to 6.4. B) Box and whisker plot shows the number of weeks until tumors reached 1 cm in diameter. (*p< 0.05). C) Box and whisker plot depicting tumor growth expressed as the number of days from when tumors are measurable (3 mm) until tumors reached 1 cm in diameter. (p=0.08).

Article Snippet: The Mouse Gas6 Quantikine® ELISA kit (R&D Systems, Minneapolis, MN, US) was used according to the manufacturer’s instructions.

Techniques: Transplantation Assay, Whisker Assay