mouse primary antibody against glial fibrillary acidic protein gfap  (Cell Signaling Technology Inc)

Journal: Bioactive Materials

Article Title: A self-locking conductive cardiac patch for immediate electrical integration with infarcted rat myocardium

doi: 10.1016/j.bioactmat.2025.10.045

Figure Lengend Snippet: The effects of patches on angiogenesis and inflammation in infarcted tissues 4 weeks post-MI. a) Immunofluorescence staining of α-smooth muscle actin (α-SMA, a vascular protein marker, green) and von Willebrand factor (vWF, an endothelial marker, red) of the infarct region in the MI, BMN-P, BMN-CP and CBMN-CP groups. b, c) Quantitative analysis of α-SMA (b) and vWF (c) in different groups based on fluorescent staining images (n = 4). d) Representative immunofluorescence staining of the infarct region in different groups for CD86 (green) and CD206 (red). e, f) Fluorescence intensity statistics of CD86 (e) and CD206 (f) in different groups based on fluorescent staining images (n = 4). Nuclei are stained blue with DAPI. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: The tissue sections were then incubated with the following primary antibodies: rabbit Connexin 43 primary antibody (BOSTER, BA1727, 1:200, China), mouse α-actinin primary antibody (Abcam, AB9465, 1:200, UK), mouse α-SMA primary antibody (Wuhan Sanying, 67735-1-IG, 1:400, China), rabbit vWF primary antibody (Wuhan Sanying, 27186-1-AP, 1:300, China), mouse CD86 primary antibody (BOSTER, BA4121, 1:100, China) and rabbit CD206 primary antibody (Wuhan Sanying, 18704-1-AP, 1:400, China).

Techniques: Immunofluorescence, Staining, Marker, Fluorescence

Journal: bioRxiv

Article Title: Comparative profiling of SARS-CoV-2 variant infections reveals diverse impacts on host cell RNA and RNA binding protein distribution and regulation

doi: 10.64898/2026.01.22.701202

Figure Lengend Snippet: A) Architecture of the SARS-CoV-2 genome, with black lines indicating canonical junctions yielding subgenomic RNAs. Mutations in pre-VOC, Alpha, Delta and Omicron BA.1 isolates are shown relative to the SARS-CoV-2 reference sequence (NC_045512.2). Non-coding, synonymous and non-synonymous mutations are highlighted in grey, black and red, respectively. B) Representative flow-cytometry plots showing nucleocapsid intensity at 48 hpi. C) Immunofluorescence staining of infected Calu-3 cells with an antibody against SARS-COV-2 nucleocapsid and spike proteins. Scale bars: 40 µm. D) Immunofluorescence staining for nucleocapsid protein combined with viral RNA FISH in pre-VOC, Omicron BA.1 and additional Omicron isolates corresponding to BA.1, BA.5 and JN.1 subvariants. E) Representative bioanalyzer traces showing similar levels of 18S and 28S rRNA between infected and mock samples.

Article Snippet: Samples were immunolabeled with an anti-nucleocapsid primary antibody (Sinobiological, 40143-MM05-100) and an Alexa Fluor 647-conjugated secondary antibody as previously described After.

Techniques: Sequencing, Flow Cytometry, Immunofluorescence, Staining, Infection

Journal: bioRxiv

Article Title: Comparative profiling of SARS-CoV-2 variant infections reveals diverse impacts on host cell RNA and RNA binding protein distribution and regulation

doi: 10.64898/2026.01.22.701202

Figure Lengend Snippet: A) Schematic of the experimental workflow used in this study. B) Architecture of the SARS-CoV-2 genome, with binding sites of FISH probe sets indicated by color triangles. C) Immunofluorescence staining of infected Calu-3 cells with an antibody against SARS-CoV-2 nucleocapsid protein (N-protein) combined with viral RNA smiFISH. Scale bar: 40 µm. D) Mean of total viral RNA FISH and nucleocapsid IF intensities per infected cell, as shown performed in ( C ), with standard deviation across n=3 replicates . E) Viral RNA smiFISH conducted on control or infected Calu-3 cells using the indicated probes specific targeting the ORF1ab (green) or ORF-N (magenta) sub-genomic regions. Scale bar: 20 µm. F) Location of RT-qPCR primers used to assess ORF1ab and ORF N sub-genomic RNA expressions and corresponding RT-qPCR 2 -ΔΔCt results using pre-VOC as the reference. Data represents n=3 replicates. Statistical significance is denoted as follow: * p<0.05, ** p<0.005, *** p<0.0005. G) Viral RNA smiFISH targeting positive-sense (+) and negative-sense (-) viral RNA, combined with immunofluorescence co-labeling with the 9D5 antibody detecting dsRNA structures. Scale bar: 10 µm.

Article Snippet: Samples were immunolabeled with an anti-nucleocapsid primary antibody (Sinobiological, 40143-MM05-100) and an Alexa Fluor 647-conjugated secondary antibody as previously described After.

Techniques: Binding Assay, Immunofluorescence, Staining, Infection, Standard Deviation, Control, Quantitative RT-PCR, Labeling

Journal: bioRxiv

Article Title: Comparative profiling of SARS-CoV-2 variant infections reveals diverse impacts on host cell RNA and RNA binding protein distribution and regulation

doi: 10.64898/2026.01.22.701202

Figure Lengend Snippet: A) Number of total proteins (A) or ISG (B) detected across Mock and infected conditions. Proteins significantly up- and downregulated following infection (padj < 0.05, log2-fold change > 1 or < −1) are highlighted in red and blue, respectively. B) Two-dimensional kernel density estimates of RNA log2 fold changes versus protein log2 fold changes for each condition (solid contours; p < 0.05). The density maximum for each condition is indicated by a cross. Dashed lines represent linear regressions, and the corresponding Pearson correlation coefficients (R) are annotated. C) Number of genes binned by their RNA and protein relative expression for each variant. ‘up’ and ‘down’: log2 fold-change > 1 and < −1, respectively, and adjusted p-value < 0.05. nc: not significantly changed. Numbers indicate the gene count per bin. Bubble sizes represent the percentage of differentially expressed genes, either at the RNA of protein level. Colors encode the combined direction of RNA and protein expression change (concordant up, concordant down or discordant changes). D) Heatmap showing the standardized degradation rate of proteins upregulated (up, padj<0.05 and fold-change>1.2), not changed (nc) or downregulated (down, padj<0.05 and fold-change<0.8) across infections, and the corresponding number of proteins. E) Clustermap of standardized, batch-corrected protein intensities across individual replicates (N=4) and conditions for each SRP component. F) Immunofluorescence staining of IRF2 and N-protein in uninfected Calu-3 cells (mock) or in cells infected with pre-VOC or Omicron viruses. G) Fluorescent OPP staining of infected cells combined with nucleocapsid immunofluorescence. OPP(-): negative control without OPP. OPP(+): mock cells with OPP. Scale bar represents 20 µm.

Article Snippet: Samples were immunolabeled with an anti-nucleocapsid primary antibody (Sinobiological, 40143-MM05-100) and an Alexa Fluor 647-conjugated secondary antibody as previously described After.

Techniques: Infection, Expressing, Variant Assay, Immunofluorescence, Staining, Negative Control

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A-B. Group mean +/- SEM percent of fluorescent radiant efficiency (p/s)/(µW/cm 2 ) relative to radiant efficiency at 24 hr timepoint measured at site of injection for 50-, 100- or 200-cycle ALD coated powders administered at 2 mg/mL or (B) 0.4 mg/mL. Unconjugated, liquid IVISense680 fluorescent dye, conjugated OVA-IVISense680 or Alhydrogel-adsorbed, conjugated OVA-IVISense680 were administered at equimolar concentrations to amount of fluorescent dye in ALD coated powder at 2 mg/mL dose. Non-linear 4PL regression fit line with 95% confidence intervals shown for ALD coated powders (constraints on 4PL regression bottom = 0, top = 100) C. Correlation of time to 50% particle dissolution in vitro and time to 50% fluorescent signal loss in vivo (using 4PL fit parameters) at indicated doses. Simple linear regression of data at each dose shown, with slope of regression line indicated on plot. D-E. Anti-OVA IgG1 seroconversion percentage following vaccination with 100-cycle ALTA ® OVA dosed at 2 mg/mL (960 ng OVA) or 0.4 mg/mL (192 ng OVA) or (E) 200-cycle ALTA ® OVA dosed at 2 mg/mL (920 ng OVA) or 0.4 mg/mL (192 ng OVA). The IgG1 data is overlaid with group mean +/- SEM percent of fluorescent radiant efficiency relative to 24 hr timepoint measured at site of injection. Dotted lines at week 2 or week 7 indicate shift in rate of fluorescent signal loss for (D) 100-cycle or (E) 200-cycle powders, respectively.

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection, Dissolution, In Vitro, In Vivo

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A. Study design, n=10 mice per group. All mice received 62.5 ng OVA dose, either in a single injection, or over the course of 7 daily injections. Of note, Al 3+ ion content differs significantly between groups treated with ALD-coated material or Alhydrogel. B. Anti-OVA IgG1 titers at week 3 post first injection (14 days post final injection for 7 day dose groups). Plot shows geometric mean titer (n=10/group) with 95% confidence interval, **** = p < 0.0001. Data analyzed using Kruskal-Wallis test. C. Anti-OVA IgG1 titers throughout study, plot shows geometric mean titer (n=10/group) with 95% confidence interval. D. Anti-OVA IgG1 seroconversion percentage, indicating a 2 log-fold increase in anti-OVA IgG1 titers relative to pre-injection baseline.

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A. Anti-OVA IgG1 titers following administration of 200 ng OVA dose given from Alhydrogel-adsorbed OVA liquid prime/boost (injection schedule 100 ng D0/100 ng D28 or 100 ng D0/100 ng D49). For mixed products (green/purple traces) 100- or 200-cycle ALTA ® powders containing a 100 ng OVA dose were resuspended in a diluent containing 100 ng Alhydrogel-adsorbed OVA and given as a single injection on D0. Plots shows geometric mean titer (n=8-10/group) with 95% confidence interval. B. Total AUC of log10-transformed anti-OVA IgG1 titers shown in A from week 0 to week 16. Column shows mean AUC +/- SEM (n=8-10 mice/group) Data analyzed using one-way ANOVA with Tukey’s multiple comparisons test. **** = p < 0.0001. *** = p < 0.001. ** = p < 0.01. C. The percentage of OVA-specific CD8+ T cells relative to total CD8+ T cells in whole blood was measured using flow cytometry (see methods). Plot shows mean +/- SEM (n=5 mice/group). D. Anti-OVA IgG1 titers following a 200 ng OVA dose given in 50-, 100- or 200-cycle ALD coated vaccine powder. For the mixed ALD vaccine product (purple), 50-cycle and 200-cycle materials were resuspended in diluent at a 2x concentration, then mixed at a 1:1 ratio immediately prior to injection to deliver a total dose of 200 ng OVA. Plot shows geometric mean titer (n=9-10/group) with 95% confidence interval. E. Total area under the curve of anti-OVA IgG1 titer was calculated for all animals after vaccination with 200 ng OVA from 50-, 100-, 200- or mixed 50+200-cycle ALTA ® OVA powders. Plot shows the group mean AUC +/- SEM (n=9-10/group). Data analyzed using one-way ANOVA with Tukey’s multiple comparisons test. * = p < 0.05. F. The percentage of OVA-specific CD8+ T cells relative to total CD8+ T cells in whole blood was measured using flow cytometry (see methods). Plot shows mean +/- SEM (n=5 mice/group).

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection, Transformation Assay, Flow Cytometry, Concentration Assay

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A. Total anti-N332-GT5 gp140 IgG titers after a single administration of 50-cycle ALTA ® on D0 at indicated doses of N332-GT5 gp140. Plots shows geometric mean titer (n=8/group) +/- 95% confidence interval. B. Analysis of fluorescent signal at site of injection following administration of 50-cycle ALTA ® containing fluorescently-labeled N332-GT5 gp140. Plot shows mean +/- SEM of percent of fluorescent radiant efficiency (p/s)/(µW/cm 2 ) relative to radiant efficiency at 24 hr timepoint measured at site of injection for 50-cycle ALTA ® N332-GT5 gp140 administered at specified doses (square traces). Total anti-N332-GT5 gp140 IgG titers plotted as geometric mean +/- 95% confidence interval (n=5 mice/group) (triangle traces). C. Kinetics of total anti-N332-GT5 gp140 IgG titers elicited by a single administration of 10 µg N332-GT5, delivered in a liquid formulation (blue) or in 50-cycle ALTA ® products with/without the presence of adjuvants in the injection diluent. Plot shows geometric mean titer +/- 95% confidence interval (n=16 mice/group red trace, n=8 mice/group all others) D. Total anti-N332-GT5 gp140 IgG1, IgG2b, IgG2c and IgG3 titers elicited by a single administration of 10 µg N332-GT5 gp140 at week 8 post injection, delivered in a liquid formulation (blue) or in 50-cycle ALTA ® products with/without the presence of adjuvants in the injection diluent. Plot shows geometric mean titer +/- 95% confidence interval (n=16 mice/group red trace, n=8 mice/group all others). ** = p < 0.01, * = p < 0.05, data analyzed using Kruskal-Wallis test. E-H. Total anti-N332-GT5 gp140 (E) IgG1, (F) IgG2b, (G) IgG2c and (H) IgG3 titers elicited by a single administration of 10 µg N332-GT5 gp140 at week 8 post injection, delivered in a liquid formulation (blue) or in 50-cycle ALTA ® products with/without the presence of adjuvants in the injection diluent. Plot shows geometric mean titer +/- 95% confidence interval (n=16 mice/group red trace, n=8 mice/group all others. **** = p < 0.0001, ** = p < 0.01, * = p < 0.05, ns = p > 0.05, data analyzed using Kruskal-Wallis test.

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection, Labeling, Formulation

Journal: Neural Regeneration Research

Article Title: Positive impact of indicaxanthin from Opuntia ficus-indica fruit on high-fat diet–induced neuronal damage and gut microbiota dysbiosis

doi: 10.4103/NRR.NRR-D-23-02039

Figure Lengend Snippet: Indicaxanthin ameliorates brain inflammation induced by HFD. (A) Transcription levels of iNOS, TNF-α, and IL-6 determined by real-time PCR in the brain. (B) Brain expression of COX-2 and iNOS. (C) Densitometric analysis of COX-2 and iNOS protein levels normalized to β-actin levels. (D) Brain expression of cytosolic p65 and nuclear p65. (E) Densitometric analysis of cytosolic p65 protein levels normalized to β-actin levels and nuclear p65 levels normalized to laminin B levels. (F) Representative images of GFAP-positive cells (red) on the surface. Hoechst staining was used to label the nuclei (blue) (microscope magnification 10×); scale bars: 100 μm. (G) Percentage of GFAP-positive cells. (H, I) Plasma circulating levels of TNF-α (H) and IL-1β (I). Data are presented as the mean ± SEM ( n = 8/group). *** P < 0.001, vs . STD mice; ### P < 0.001, vs. HFD-fed mice. COX-2: Cyclooxygenase-2; GFAP: glial fibrillary acidic protein; HFD: high-fat diet; IL-6: interleukin-6; Ind: indicaxanthin; iNOS: inducible nitric oxide synthase; PCR: polymerase chain reaction; STD: standard diet; TNF-α: tumor necrosis factor-α.

Article Snippet: Next, the sections were incubated with a mouse primary antibody against glial fibrillary acidic protein (GFAP) (1:300, Cell Signaling Technology, Danvers, MA, USA, Cat# 3670, RRID: AB_10693476) at 4°C overnight and then with an anti-mouse IgG-Alexa Fluor 594 secondary antibody (goat, 1:300, Invitrogen, Cat# A-11005, RRID: AB_2534073) for 2 hours at room temperature.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Staining, Microscopy, Clinical Proteomics, Polymerase Chain Reaction