mouse apoe (apolipoprotein e) elisa kit  (Elabscience Biotechnology)

Journal: Journal of Neuroinflammation

Article Title: The protective PLCγ2-P522R variant mitigates Alzheimer’s disease-associated pathologies by enhancing beneficial microglial functions

doi: 10.1186/s12974-025-03387-6

Figure Lengend Snippet: PLCγ2-P522R variant does not increase the plaque-associated APOE or DAM signature in female APP/PS1 mice. A Analysis of total APOE and APOE areas (µm 2 ) within and surrounding (within 0–10- and 10–20-µm distances from the amyloid plaque outline) β-amyloid plaques revealed no differences between the APP/PS1xPLCγ2-P522R (A+/P ki/ki ) and APP/PS1 (A+/P wt/wt ) mice. A+/P wt/wt n = 5, A+/P ki/ki n = 6. Colored disks on the x-axis indicate the analyzed area overlapping (middle circle) and surrounding β-amyloid plaques. The scale bar in the representative immunofluorescence images is 50 μm. Unpaired samples t test. All the data are presented as the means ± SEMs. Each data point represents an individual mouse. B Heatmap of z scored vst-normalized expression of homeostatic as well as stage 1 and stage 2 disease-associated microglia (DAM) signature genes in the bulk temporo-occipital cortex and C ) CD11b+ microglia isolated from the whole brain tissue of 13-month-old wild-type (A-/P wt/wt ), A+/P wt/wt and A+/P ki/ki female mice. Cortex A-/P wt/wt n = 1, A+/P wt/wt n = 4, and A+/P ki/ki n = 4; CD11b+ A-/P wt/wt n = 7, A+/P wt/wt n = 4, and A+/P. ki/ki n = 4

Article Snippet: Soluble and insoluble APOE were measured in lysates using an Apolipoprotein E (APOE) Mouse ELISA Kit (#ELK2007, Gentaur) according to the manufacturer’s instructions.

Techniques: Variant Assay, Immunofluorescence, Expressing, Isolation

Journal: Molecular Neurodegeneration

Article Title: Alzheimer’s disease associated isoforms of human CD33 distinctively modulate microglial cell responses in 5XFAD mice

doi: 10.1186/s13024-024-00734-8

Figure Lengend Snippet: Altered Aβ plaque composition by hCD33 isoforms. a Representative confocal fluorescent images showing the different degrees of plaque compaction by co-staining for total Aβ and ThioS. A diffuse plaque without ThioS staining at the far left, a plaque with small ThioS core in the middle, and a highly compact plaque at the far right are presented. Scale bar = 20 µm ( b-e ). Quantification of Aβ deposits containing a ThioS. + core. Representative epifluorescent images of Aβ deposits in the dorsal subiculum of 5XFAD mice at ( b ) 4 and ( d ) 8 months, with co-staining with anti-Aβ antibody (white) and ThioS (blue). Scale bar = 50 µm. Quantification for pooled males (squares; n = 5 per genotype) and female (circles; n = 5/genotype) mice at ( c ) 4 and ( e ) 8 months. f Representative confocal fluorescent images of plaque composition in the dorsal subiculum of 8 months old CD33M and CD33m mice. Scale bar = 20 µm ( g , h ) Quantification of plaque composition (ratio of Aβ over ThioS levels) in the dorsal subiculum of 5XFAD mice at ( g ) 4 and ( h ) 8 months. A total of 300 plaques per mouse, from 5 males and 5 females, were quantified for each group. i Biochemical characterization of PK-sensitivity of insoluble Aβ 1-42 from 5XFAD mice. j Quantification of the percentage of insoluble Aβ 1-42 levels after PK digestion compared to not treated with PK. k Quantification of the ratio of ApoE to Aβ 1-42 in the insoluble fraction from 5XFAD mice ( n = 9, 10, and 9 mice of control, CD33M, and CD33m genotypes, respectively)

Article Snippet: ApoE levels were quantified in both the soluble and insoluble fractions using a Mouse Apolipoprotein E (APOE) ELISA Kit (Mybiosource).

Techniques: Staining, Control

Journal: Molecular Neurodegeneration

Article Title: Alzheimer’s disease associated isoforms of human CD33 distinctively modulate microglial cell responses in 5XFAD mice

doi: 10.1186/s13024-024-00734-8

Figure Lengend Snippet: Single-cell RNA sequencing reveals Ccl3 and Trem2 DAM enriched in the CD33m genotype. a Unsupervised and iterative machine-learning based clustering of 15,200 microglia ( Hexb + Fcrls + Tmem119 + Sall1 + ) and BAM ( Ms4a7 + Mrc1 + Lyve1 + Timd4 + ) collected from 5XFAD control, CD33M 5XFAD, CD33m 5XFAD, and control non-5XFAD mice. Microglia from three homeostatic (HM1-3), two transitioning (TM1-2), two RNA binding protein (RBM1-2) subpopulations along with the disease-enriched interferon responsive (IRM), myelin transcript enriched (MTEM), and disease associated (DAM) clusters. b-e UMAPs of individual samples showing ( b ) 5XFAD control, c CD33m 5XFAD, d CD33M 5XFAD, and e control non-5xFAD. f , g Separation of each cluster by ( f ) proportion of cells and ( g ) absolute number of cells belonging to each genotype. DAM, specifically Trem2 DAM, are enriched in the CD33m group and reduced in the CD33M group. h Differential gene expression per cluster used to define microglial subpopulations. HM are characterized by homeostatic genes ( P2ry12, Tmem119 ), RBM by genes related to RNA binding ( Son, Fus ), TM by a combination of homeostatic ( P2ry12, Tmem119 ), complement ( C1qa, C1qb ), and proliferative ( MKi-67, Top2a ) genes. DAM are defined by Clec7a and Apoe expression and further delineated by expression of Ccl3 and Trem2 . i UMAP showing enriched Nes expression within the Ccl3 DAM cluster. j-l Violin plots showing the upregulation of ( j ) Nes , k Jun , and l Fos in the CD33m + 5XFAD group relative to 5XFAD control and CD33M + 5XFAD

Article Snippet: ApoE levels were quantified in both the soluble and insoluble fractions using a Mouse Apolipoprotein E (APOE) ELISA Kit (Mybiosource).

Techniques: RNA Sequencing, Control, RNA Binding Assay, Gene Expression, Expressing

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: ApoE is the most highly expressed immune suppressive gene in the melanoma B16-F10 cell lines and apoE serum levels rise with tumor growth in vivo. (A) mRNA expression as revealed by nanostring nsolver Pancancer immune profiling, of the immune suppressive marker genes in B16F10 cells (n=6). (B) ApoE expression was undetectable in the serum of apoE knockout (apoE-/-) mice, but present at high levels in wild type (C57/BL6) mice with ELISA assay. (C) ApoE knockout mice were inoculated with 10^4 wild type (B16F10) cells and serum levels of apoE increased over time with increasing tumor size. (D) Validation of apoE gene knockout in B16F10 cells by CRISPR-Cas9 gene deletion. The level of apoE protein expression was measured in WT B16F10 and the corresponding apoE-/- clone by Western Blot. (E) Equivalent numbers of WT and apoE-/- B16 cells were plated and apoE levels released into culture media were quantified by ELISA at 48hr. ApoE is secreted at high levels in WT B16 cells and is not detectable in the apoE-/- cell line. (F) To evaluate whether targeting apoE influenced cell viability and proliferation, 5x10^4 WT (n=6) or apoE-/- B16 cells (n=6) were grown in 12-well plates and proliferation rates were measured at 24hr and 48hr by MTT assay. There is no statistically significant difference in cell proliferation rate between the two groups. (G) Cell cycle distribution was determined in WT and apoE-/- B16 cells. The various phases of the cell cycle are differentiated in the flow cytometry plot on the left: G0-G1 is the pre-synthesis phase, S-phase cells are undergoing active DNA synthesis and G2-M cells are preparing for mitosis. Bar graphs represents the percentage of cells in G0-G1, S and M phase of the cell cycle. Cell counts and cell cycle distribution indicate that WT and apoE-/- B16 cells proliferate at equivalent rates. Data are representative of three independent experiments. Results are expressed as mean score ±SD. ***p<0.001, determined by unpaired two-tailed Student’s t-test.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: In Vivo, Expressing, Marker, Knock-Out, Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Gene Knockout, CRISPR, Western Blot, MTT Assay, Flow Cytometry, DNA Synthesis, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: ApoE secreted into media from B16 melanoma tumor cells inhibits T-cell proliferation and function. (A) Conditioned media (CM) from WT or apoE -/- cells was cultured with T-cells activated by CD3/CD28 beads. Bar graphs depict IFNγ released under these conditions. The production of IFNγ was significantly suppressed when T cells were cultured in WT B16 CM, but this suppression was reversed in apoE free CM from apoE -/- cells. (B) Representative flow cytometry plots and (C) quantification of T cell apoptosis in CM. Results show that T cell viability is markedly reduced in the WT CM, whereas induction of apoptosis was reversed when cells were cultured in the apoE -/- medium, similar to RPMI control medium alone. (D) Representative flow cytometry plots and (E) quantification of cell cycle analysis of T cells in different media. Cell cycle distribution showed an arrest of activated T cells in the G0-G1 phase in the WT B16 medium, while in the apoE -/- medium most T cells were distributed in the S and G2-M phases, similar to the RPMI media control. Results are expressed as mean score ± SD. *p<0.05; **p<0.005; ***p<0.001, determined by unpaired two-tailed Student’s t-test.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Cell Culture, Flow Cytometry, Control, Cell Cycle Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: ApoE agonist peptide COG133 inhibits cytokine secretion induced by immunogenic tumor cells, while anti-APOE blocking antibody enhances cytokine secretion in tumor cell/splenocyte reactions. (A) Naïve and vaccinated splenocyte production of IFNγ was markedly reduced when splenocytes were co-cultured with immunogenic (BET/JQ1 treated) B16 tumor cells in the presence of the apoE agonist COG133. (B) Naïve splenocyte production of IFNγ was significantly increased when co-cultured with immunogenic B16 tumor cells with anti-APOE antibody. (C) . COG133 was also inhibitory of IFNγ production when vaccinated splenocytes were used for co-culture with treated immunogenic cancer cells. It is of interest to note that the levels of IFNγ production is significantly greater when vaccinated splenocytes were used for this experiment. NS: naïve splenocytes, splenocytes were collected from naïve C57BL/6 mice. Treated B16: B16 cells were expose to Myc inhibitor (0.25µM BET and 0.25µM JQ1) for 4 days, to induce immunogenicity. VS: vaccinated splenocytes. Irradiated 10 4 WT B16 tumor cells and 100ug/ml anti-CTLA4 antibody were administered to C57BL/6 mice on day 0, and splenocytes were collected at day 7 after tumor cell inoculation. Data are representative of three independent experiments. Results are expressed as mean score ± SD. *p<0.05; **p<0.005; ***p<0.001, determined by unpaired two-tailed Student’s t-test.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Blocking Assay, Cell Culture, Co-Culture Assay, Immunopeptidomics, Irradiation, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: lrp8 is the most dominant receptor expressed on activated T cells and blocking lrp8 enhanced T-cell activation. (A) Expression of apoE receptors shows predominance of ldlr and lrp8 receptors on T cells, (B) lrp8, ldl and lrp1 receptors on dendritic cells, and (C) lrp1 receptors on macrophages as shown by quantitative real-time PCR (qPCR). Expression of ldlr and lrp8 was upregulated following T cell stimulation with CD3/CD28 beads. Expression of lrp8 was also upregulated following dendritic cell stimulation with TLR7/8. (D) The amount of IFNγ production from vaccinated splenocytes co-cultured with immunogenic WT B16 cells was quantified by ELISA assay. Results show that IFNγ production was significantly suppressed in the presence of the apoE agonist COG133, but suppression did not occur when splenocytes were harvested from lrp8 -/- mice. These findings suggest that T-cell function is at least partially inhibited by apoE through the lrp8 receptor pathway. Results are expressed as mean score ± SD. *p<0.05; **p<0.005; ***p<0.001, determined by unpaired two-tailed Student’s t-test.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Blocking Assay, Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Cell Stimulation, Cell Culture, Enzyme-linked Immunosorbent Assay, Cell Function Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: Knock out of apoE in both B16 tumor cells and in mice enhances host immunity and attenuates tumorigenicity. (A) In vivo, 104 WT or apoE-/- B16 cells were injected into the right leg of WT (n=9), apoE-/- (n=9) or lrp8-/- mice (n=9). The average tumor growth in each group (n=6) is compared. Tumor growth was significantly slower when apoE-/- B16 cells were injected into apoE-/- or lrp8-/- mice versus the other groups (two-way ANOVA; P <0.0001). (B) The final tumor volume between treatment groups at the end point of the experiment (Day 20) was also compared using a one-way ANOVA followed by Tukey HSD pairwise multiple comparisons between treatment groups. Tumor volumes at the end point of the experiment were significantly different between treatment groups (one-way ANOVA; P <0.0001). *p<0.05; **p<0.01; ****p<0.0001.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Knock-Out, In Vivo, Injection

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: Survival curve (A) Survival was significantly better (n=9) when apoE -/- B16 cells were injected into apoE -/- or lrp8 -/- mice versus the other groups. (B) The median survival time and the cumulative survival probability were calculated and compared using the Kaplan-Meier survival estimator followed by a log-rank test, and the hazard ratio (HR) was calculated using the Cox proportional-hazards regression model. The comparison between the groups is shown in the table.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Injection, Comparison

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: The first three tumors that reached 15mm in each of the experimental groups were harvested and the immune cell infiltrate and immune pathway activation was analyzed with nanostring Pancancer immune profiling. (A) Immune cell type scores and (B) immune pathway (signature) scores show that immune cell infiltrates and immune pathway activation were greatest when apoE-/- cells were injected into apoE-/- mice. Dot line plots show the score trends of 12 immune cell lines and 29 immune pathways. Box plots show selective representative score comparisons. Results are expressed as mean score ±SD. *p<0.05, determined by unpaired two-tailed Student’s t-test. Wt/wt: wt B16 cells injected in wt mice; apoE-/-/apoE-/-: apoE-/- cells injected in apoE-/- mice; apoE-/-/lrp8-/-: apoE-/- cells injected in lrp8-/- mice; apoE-/-/wt: apoE-/- cells injected in wt mice; wt/apoE-/-: wt cells injected in apoE-/- mice; wt//lrp8-/-: wt cells injected in lrp8-/- mice.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Activation Assay, Injection, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: Nanostring PanCancer Immune Profiling analysis of RNAseq of activation markers for T cells and DCs that infiltrated into the tumors from the 6 different groups. (A) Activated T cell genes and (B) activated DC genes were all statistically significantly increased in the tumor from apoE-/-/apoE-/- group when compared with wt/wt control groups. The relative gene expression level was indicated on the y-axis and tumor groups are listed along the x-axis. Results are expressed as mean score ±SE. *p<0.05, determined by unpaired two-tailed Student’s t-test.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Activation Assay, Control, Gene Expression, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: To validate nanostring results, both CD45 and CD3 expression were examined with IHC staining in the tumors from WT or apoE-/- mice following injection with WT or apoE-/- tumor cells. Representative images of CD45 (A) and CD3 (C) staining visualized with DAB (brown) and counterstained with hematoxylin (blue, nuclei). (B, D) Optical density (mean gray value) obtained by color deconvolution analysis. Optical density graph bars represent the mean ± SD (n = 30 images). ***p<0.001, determined by unpaired two-tailed Student’s t-test. Wt/wt: wt B16 cells injected in wt mice; apoE-/- /apoE-/-: apoE-/- cells injected in apoE-/- mice.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Expressing, Immunohistochemistry, Injection, Staining, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: ApoE knock out in B16 tumor cells induces potent immunogenicity. Wild type and apoE -/- mice (A) or wild type and lrp8 -/- mice (B) , were inoculated with either WT B16 or apoE -/- cells, with anti-CTLA4 antibody on day 0. Splenocytes were harvested on day 7 and co-cultured with either WT B16 or apoE -/- B16 cells for 48hr, following which IFNγ levels in media were compared with ELISA assay. Results show that IFNγ production is highest for all groups when apoE -/- cells are used. These findings re-iterate the potent inhibitory effect of apoE on immune cell activation in the cancer environment. Results are expressed as mean score ± SD. *p<0.05; **p<0.005; ***p<0.001, determined by unpaired two-tailed Student’s t-test.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Knock-Out, Immunopeptidomics, Cell Culture, Enzyme-linked Immunosorbent Assay, Activation Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: ApoE RNA-seq expression is abundant in cutaneous melanoma but is not associated with PD-L1 or PD1. Scatterplots showing mRNA expression correlation of APOE (x-axis) with PD-L1, PD1 and APOC1 from the TCGA melanoma datasets based on their RNA-seq gene expression values (measured by RSEM algorithm). APOE expression did not correlate with PD-L1 or PD1 but did positively correlate with APOC1 expression which was used as a control gene. The correlation was evaluated by the Spearman correlation coefficient with a cut-off value of 0.5 and P-value used a cut-off value of 0.05.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: RNA Sequencing, Expressing, Gene Expression, Control

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: Scatterplots show the tumor purity-corrected partial Spearman’s rho value and the correlation between gene expression with infiltration of six immune cell estimates. Top row is APOE expression while the bottom row depicts PD-L1. PD-L1 correlated with CD8 T cell and neutrophil infiltrates, while APOE does not correlate with immune cell infiltrates. The correlation was evaluated by the Spearman correlation coefficient with a cut-off value of 0.5 and P-value used a cut-off value of 0.05.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Gene Expression, Expressing

Journal: Frontiers in Immunology

Article Title: Tumor Apolipoprotein E is a key checkpoint blocking anti-tumor immunity in mouse melanoma

doi: 10.3389/fimmu.2022.991790

Figure Lengend Snippet: Cumulative survival of patients associated with bifurcate gene expression at 30%. TCGA database includes primary and metastasis samples (n=462). APOE expression did not associate with survival, while PD-L1 and PD1 was associated with survival at this level of analysis. The correlation was evaluated by the Spearman correlation coefficient with a cut-off value of 0.5 and P-value used a cut-off value of 0.05.

Article Snippet: ApoE levels in mouse sera were quantified using the mouse apoE ELISA PRO kit from Mabtech (Cincinnati, OH) as per the manufacturer’s directions.

Techniques: Gene Expression, Expressing