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mouse vegf quantikine elisa kit  (R&D Systems)
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R&D Systems mouse vegf quantikine elisa kit
T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by <t>ELISA.</t> n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures
Mouse Vegf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures

Journal: Molecular Medicine

Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

doi: 10.1186/s10020-026-01651-w

Figure Lengend Snippet: T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures

Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

Techniques: Migration, Activity Assay, Isolation, Cell Culture, Analysis, Counting, Co-culture Assay, Derivative Assay, WST Assay, Comparison, Enzyme-linked Immunosorbent Assay, Knock-Out

Regulation of cytokines and angiogenesis markers in the tumor. A OLINK proteomic analysis of inflammatory cytokines and chemokines in the tumors of wildtype and IKKε KO mice at the end of the experiment. Heat map and Volcano Plot of significantly different regulations. n = 4/group ( B ) Cxcl1 and Cxcl2 ELISA. Cxcl1: WT n = 8, IKKε-KO n = 6; Cxcl2: WT n = 6, IKKε-KO n = 5. C Determination of CD31-positive cells as vessel markers in the tumors. Left panel: FACS analysis, n = 6 per group. Right panel: immunofluorescence images and quantitative analysis, n = 4–5 tumors per group; 3–5 areas per tumor were evaluated. CD31-positive areas were normalized to wildtype controls to facilitate comparison. D MMP-9 expression as assessed by Western blot and zymography, n = 5–6 per group. E HIF-1α expression as determined by immunofluorescence, the pictures show on representative stain from 6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. F VEGF expression analyzed by ELISA and Western Blot, n = 3/group. Student´s t-test, * P < 0.05, ** P < 0.01

Journal: Molecular Medicine

Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

doi: 10.1186/s10020-026-01651-w

Figure Lengend Snippet: Regulation of cytokines and angiogenesis markers in the tumor. A OLINK proteomic analysis of inflammatory cytokines and chemokines in the tumors of wildtype and IKKε KO mice at the end of the experiment. Heat map and Volcano Plot of significantly different regulations. n = 4/group ( B ) Cxcl1 and Cxcl2 ELISA. Cxcl1: WT n = 8, IKKε-KO n = 6; Cxcl2: WT n = 6, IKKε-KO n = 5. C Determination of CD31-positive cells as vessel markers in the tumors. Left panel: FACS analysis, n = 6 per group. Right panel: immunofluorescence images and quantitative analysis, n = 4–5 tumors per group; 3–5 areas per tumor were evaluated. CD31-positive areas were normalized to wildtype controls to facilitate comparison. D MMP-9 expression as assessed by Western blot and zymography, n = 5–6 per group. E HIF-1α expression as determined by immunofluorescence, the pictures show on representative stain from 6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. F VEGF expression analyzed by ELISA and Western Blot, n = 3/group. Student´s t-test, * P < 0.05, ** P < 0.01

Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

Techniques: Analysis, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Comparison, Expressing, Western Blot, Zymography, Staining

Early regulation of cytokines and angiogenesis markers in the tumor. A Cxcl1 ELISA, ( B ) MMP-9 expression (Western Blot and zymography), ( C ) HIF-1α expression as determined by immunofluorescence. The pictures show a representative stain from 4–6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. D FACS analysis of CD31. + -cells as vessel markers in the tumors, ( n = 6/group). E VEGF expression analyzed by ELISA and Western Blot, ( F ) p65 expression and activity in nuclear extracts as determined by Western Blot and transcription factor ELISA. n = 3–5/group. Student´s t-test, * P < 0.05, ** P < 0.01

Journal: Molecular Medicine

Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

doi: 10.1186/s10020-026-01651-w

Figure Lengend Snippet: Early regulation of cytokines and angiogenesis markers in the tumor. A Cxcl1 ELISA, ( B ) MMP-9 expression (Western Blot and zymography), ( C ) HIF-1α expression as determined by immunofluorescence. The pictures show a representative stain from 4–6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. D FACS analysis of CD31. + -cells as vessel markers in the tumors, ( n = 6/group). E VEGF expression analyzed by ELISA and Western Blot, ( F ) p65 expression and activity in nuclear extracts as determined by Western Blot and transcription factor ELISA. n = 3–5/group. Student´s t-test, * P < 0.05, ** P < 0.01

Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Zymography, Immunofluorescence, Staining, Analysis, Activity Assay

Angiogenesis assay in HUVEC with and without depletion of IKKε by siRNA. A IKKε mRNA and protein expression in HUVEC cells after treatment with IKKε siRNA or a scrambled negative control as determined by PCR and Western Blot analysis, PCR: n = 3–4/group, Western Blot ( n = 2/group) ( B ) Tube formation assay, left side: microscopic pictures, right side: analysis of the total mesh area of naïve HUVEC, or HUVEC treated with NC and IKKε siRNA. n = 6–7/group. The scale bars represent 100 µm. C Spheroid sprout assay, left side: microscopic pictures, right side: analysis of cumulative sprout length (CSL) in spheroids of naïve HUVEC ( n = 15 spheroids), HUVEC treated with VEGF ( n = 38 spheroids) HUVEC-NC ( n = 25 spheroids) and HUVEC-IKKε siRNA ( n = 38 spheroids). Data were generated in 5 independent biological experiments. The scale bars represent 100 µm. One Way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Journal: Molecular Medicine

Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

doi: 10.1186/s10020-026-01651-w

Figure Lengend Snippet: Angiogenesis assay in HUVEC with and without depletion of IKKε by siRNA. A IKKε mRNA and protein expression in HUVEC cells after treatment with IKKε siRNA or a scrambled negative control as determined by PCR and Western Blot analysis, PCR: n = 3–4/group, Western Blot ( n = 2/group) ( B ) Tube formation assay, left side: microscopic pictures, right side: analysis of the total mesh area of naïve HUVEC, or HUVEC treated with NC and IKKε siRNA. n = 6–7/group. The scale bars represent 100 µm. C Spheroid sprout assay, left side: microscopic pictures, right side: analysis of cumulative sprout length (CSL) in spheroids of naïve HUVEC ( n = 15 spheroids), HUVEC treated with VEGF ( n = 38 spheroids) HUVEC-NC ( n = 25 spheroids) and HUVEC-IKKε siRNA ( n = 38 spheroids). Data were generated in 5 independent biological experiments. The scale bars represent 100 µm. One Way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

Techniques: Angiogenesis Assay, Expressing, Negative Control, Western Blot, Analysis, Tube Formation Assay, Spheroids, Generated