goat anti il 32 af3040 page 5 13 antibody  (R&D Systems)

Journal: PLoS ONE

Article Title: Microbial Dysbiosis in Colorectal Cancer (CRC) Patients

doi: 10.1371/journal.pone.0016393

Figure Lengend Snippet: Samples from the same individuals and colonic sites were submitted to DNA extraction and PCR. Interleukin 17 (IL17)-immunoreactive cells in colonic tissues were mainly located in the lamina propria in the normal tissues [A: colonic normal mucosa from a normal individual (high magnification x40 at the bottom), B: colonic normal mucosa from a patient with colon cancer (high magnification x40 at the bottom)] and infiltrated the tumour tissue in a the same individual than in B [C: IL17 imlmunoreactive cells infiltrating the tumour with high magnification x40 at the bottom & D: In this double staining IL17 and CD3, the goat anti-human IL-17 antibody was added first before staining with Naphthol/Fast (red) followed by the rabbit anti-human CD3 antibody that was revealed with DAB substrate (brown); this showed that CD3 was not the only cell producing IL-17.

Article Snippet: Briefly, the goat anti-human IL-17 antibody (diluted 1∶40) was added for 2 h, and then staining was undertaken using (Vectastain AP kit from Vector Laboratories, Burlingame, CA, USA), and revealed by Naphtol/Fast Red (Sigma-Aldrich).

Techniques: DNA Extraction, Double Staining, Staining

Journal: Nature

Article Title: Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease

doi: 10.1038/s41586-021-03734-6

Figure Lengend Snippet: a. Enumeration, by flow cytometry, of microglia in WT, 5XFAD, Il3−/−5XFAD mice (n=10 5xFAD mice; n=8 Il3−/−5XFAD mice). b. Multidimentional scaling (MDS) plot of RNAseq data from 5 month old 5XFAD and Il3−/−5XFAD mice (n=12, each data point represents 4 pooled mice of 2M/2F). c. Volcano plot indicating differentially regulated genes (FC>1.6, FDR<0.1, p<0.005) in microglia of 5XFAD and II3−/−5XFAD mice. d. Heatmap of key differentially regulated genes (FC>1.6, FDR<0.1, p<0.005), except Il3rɑ which is not significantly different between 5xFAD and Il3−/− 5xFAD microglia (n=12, each data point represents 4 pooled mice of 2M/2F). e. Pathway analysis of significantly regulated genes. f. Representative immunofluorescence images of microglia from 5XFAD and Il3−/−5XFAD mice along with skeletal analysis of microglia morphology (n=5 5xFAD mice; n=4 Il3−/−5XFAD mice). g. Representative immunofluorescence images of cortex sections from 5XFAD and Il3−/−5XFAD mice stained for Aβ (6E10), Iba1 and DAPI and quantification of the number of microglia within 25μm of Aβ plaques (n=5 5xFAD mice; n=7 Il3−/−5XFAD mice). h. Schematic depicting segmentation and spatial analysis along with computed density gradient and diffusion rate of microglia surrounding Aβ (n=5 5xFAD mice; n=7 Il3−/−5XFAD mice). i. Three dimensional confocal images of mouse cortex (634x250x634μm) stained for Aβ (6E10) and microglia (Iba1). All mouse data are of groups of 5 month old animals of evenly mixed sex. j. Scheme of 3D human AD triculture microfluidic system where human neuron ReNcell VM progenitor derived astrocytes and neuronal cells, with or without the K670N/M671L (Swedish) and V717I (London) familial AD mutations resulting overproduction of Aβ and neurofibrillary tangle (NFT) p-tau (AD), are plated in the central chamber while human iPS-derived microglia labeled with CellTracker are plated in side chambers. k. Confocal images demonstrated p-tau (PHF1) localization with neurons (Tuj1) and the presence of astrocytes (GFAP) in the central chamber and microglia (P2RY12) in the side chamber. l. ELISA quantification of Aβ40, Aβ38, and Aβ42 in media of central chamber in 3D microfluidic system plated with control or AD cells (n=3 per group). m. Confocal imaging of IL-3 localization to GFAP+ human astrocytes and IL-3Rɑ localization to CellTracker labeled human iPS-derived microglia in 3D microfluidics system. n. Flow cytometry quantification of iPS-derived microglia that have migrated to the central chamber of the 3D microfluidics system with or without addition of human recombinant IL-3 (n=3 per group). o. Representative confocal images and image quantification of iPS microglia migration to central chamber (n=3 per group for astrocytes+neurons and AD astrocytes+neurons; n=4 AD astrocytes+neurons+rlL3). p. CCL2 and CCL4 levels in media of 3D microfluidics system (n=3 per group). *p<0.05, **p<0.01, ***p<0.001. Error bars indicate mean ± SEM.

Article Snippet: Biotinylated goat anti-rabbit IgG and horse anti-mouse IgG secondary antibodies were applied for IL-3 and IL-3Rɑ respectively (1:100, BA-1000 and BA-2000, Vector Laboratories) followed by streptavidin DyLight 594 (1:600, SA-5594, Vector Laboratories).

Techniques: Flow Cytometry, Immunofluorescence, Staining, Diffusion-based Assay, Derivative Assay, Labeling, Enzyme-linked Immunosorbent Assay, Imaging, Recombinant, Migration

Journal: Nature

Article Title: Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease

doi: 10.1038/s41586-021-03734-6

Figure Lengend Snippet: a. Il3 expression in astrocytes sorted from WT and 5xFAD mice at 5 and 12 months of age. b. Tissue IL-3 levels in various brain regions in WT mice at 4 months of age (n=4). c. Scheme of daily i.p. LPS injection into WT mice over 4 days and qPCR analysis of complement C3 and gfap in sorted astrocytes after LPS injection (n=6). d. Cerebrospinal fluid IL-3 levels (n=6-8). e. Representative images of GFAP+ astrocytes in the cortex of WT, Il3−/−, 5xFAD, and Il3−/−5xFAD mice. f. Representative images of Aβ deposits (6E10) and astrocytes (GFAP) in 5xFAD and Il3−/−5XFAD mice. g. Proportion of IL-3Rɑ+ microglia in the brain of WT mice at various ages (n=4). h. Il3rα transcript expression in brain homogenate of WT mice at various ages (n=4). i. Proportion of IL-3Rɑ+ macrophages in heart, liver, lung (interstitial and alveolar), and brain of WT and 5xFAD mice at 8 months of age (n=4). *p<0.05, **p<0.001. Error bars indicate mean ± SEM.

Article Snippet: Biotinylated goat anti-rabbit IgG and horse anti-mouse IgG secondary antibodies were applied for IL-3 and IL-3Rɑ respectively (1:100, BA-1000 and BA-2000, Vector Laboratories) followed by streptavidin DyLight 594 (1:600, SA-5594, Vector Laboratories).

Techniques: Expressing, Injection

Journal: Nature

Article Title: Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease

doi: 10.1038/s41586-021-03734-6

Figure Lengend Snippet: a. IL-3 levels in the plasma (PL) and cerebrospinal fluid (CSF) of WT and 5xFAD mice at 5 and 12 months (m.) of age (n=4 WT and 5xFAD 5m.plasma; n=14 WT 5m. CSF; n=6 5xFAD 5m. CSF; n=5 WT 12m. CSF; n=4 5xFAD 12m. CSF). b. Flow cytometry analysis of the brain of WT, Il3GFPfl/fl, and Il3GFPfl/fl5xFAD reporter mice. c. Quantification of IL3+ (GFP+) astrocytes in the brain of Il3GFPfl/fl and l3GFPfl/fl5xFAD mice (n=4 mice per group). d. Il3 mRNA transcript expression in Aldh1l1-GFP+ astrocytes, CD11b+CD45mid microglia, and CD45− cells sorted from Aldh1/1GFP mice (n=7 mice per group). e. Representative immunofluorescence images showing co-localization of IL-3-GFP+ with GFAP+ astrocytes in Il3GFPfl/fl mice. f. Quantification of IL3+ astrocytes (Aldh1/1-GFP+) in various brain regions and representative immunofluorescence images from numerous regions showing co-localization of IL-3 with GFP+ astrocytes in Aldh1/1GFP mice (n=4 per group). g. Flow cytometry and (h) analysis of IL-3Rɑ+ microglia in the brain of WT and 5xFAD mice (n=5 5m. WT mice; n=11 5m. 5XFAD mice; n=3 8m. WT mice; n=5 8m. 5XFAD mice). i. Il3ra mRNA expression in sorted microglia (n=5 5m. WT mice; n=4 5m. and 8m. 5xFAD mice; n=3 8m. WT mice). j. STAT5 phosphorylation in ex-vivo microglia simulated with recombinant IL-3. k. Representative immunofluorescence images showing co-localization of IL-3Rɑ with Iba1 in the cortex of 5xFAD mice. l. Il3rɑ transcript expression in sorted microglia from WT, Trem2−/−, 5xFAD, and Trem2−/−5xFAD mice displayed as Log2 fold change (FC) for indicated comparisons (for 4 month old mice p=3.45e−27 for 5xFAD vs WT, p=3.4e−06 for Trem2−/−5xFAD vs WT, p=1.82e−08 for Trem2−/−5xFAD vs 5xFAD; for 8 month old mice p=1.12e−93 for 5xFAD vs WT, p=1.4e−13 for Trem2−/−5xFAD vs WT, p=1.6e−40 for Trem2−/−5xFAD vs 5xFAD; FDR<0.0002; for WT n=13M/14F at 4 m., n=8M/8F at 8m.; for Trem2−/− n=11M/12F at 4m., 2M/2F at 8m.; for 5xFAD n=14M/14F at 4 m., 10M/9F at 8m.; and for Trem2−/−5xFAD n=11M/11F at 4m., 9M/8F at 8m.). ns=not significant. m. Flow cytometry analysis of IL3Rɑ+ on microglia from 5xFAD and Trem2−/−5xFAD mice (n=4 mice per group). Groups are of evenly mixed sex. *p<0.05, **p<0.01, ***p<0.001. Error bars indicate mean ± SEM.

Article Snippet: Biotinylated goat anti-rabbit IgG and horse anti-mouse IgG secondary antibodies were applied for IL-3 and IL-3Rɑ respectively (1:100, BA-1000 and BA-2000, Vector Laboratories) followed by streptavidin DyLight 594 (1:600, SA-5594, Vector Laboratories).

Techniques: Flow Cytometry, Expressing, Immunofluorescence, Ex Vivo, Recombinant

Journal: Nature

Article Title: Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease

doi: 10.1038/s41586-021-03734-6

Figure Lengend Snippet: a. Recombinant IL-3 or PBS was delivered into the cortex of 5xFAD mice. Three days later microglia localization to Aβ aggregates was assessed (n=6-7). b. Scheme of recombinant IL3 delivery intraperitoneally twice a week for 10 weeks to 5xFAD mice. c. Prior to sacrifice Ymaze behavioral testing was performed and time in the new arm was quantified (n=7-8). d. The amount of Aβ in the cortex of mice was quantified by analyzing histological sections (n=6). Groups of mice are of evenly mixed sex. **p<0.01. Error bars indicate mean ± SEM. e. Model of IL-3’s role in AD. Astrocytes produce IL-3. In response to Aβ, TREM2 signaling increases microglia IL-3Rɑ, rendering microglia responsive to astrocyte-derived IL-3. IL-3 signaling instigates microglia transcriptional and functional reprogramming leading to a signature of immune regulation, motility, and migration. IL-3-dependent reprogramming promotes clustering of microglia around Aβ enabling Aβ clearance and mitigation of AD pathology.

Article Snippet: Biotinylated goat anti-rabbit IgG and horse anti-mouse IgG secondary antibodies were applied for IL-3 and IL-3Rɑ respectively (1:100, BA-1000 and BA-2000, Vector Laboratories) followed by streptavidin DyLight 594 (1:600, SA-5594, Vector Laboratories).

Techniques: Recombinant, Derivative Assay, Functional Assay, Migration

Journal: Nature

Article Title: Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease

doi: 10.1038/s41586-021-03734-6

Figure Lengend Snippet: a. Gating strategy used to identify cell populations in the brain of all mice except Aldh1/1GFP mice. b. Backgating of GFP+ astrocytes in Il3GFPfl/fl mice. c. Gating Strategy Used to identify cell populations in the brain of Aldh1/1GFP mice. d. Gating and isotype control plots for microglia IL-3Rɑ staining. e. Representative images of IgG control antibody staining and IL-3 staining in the brain.

Article Snippet: Biotinylated goat anti-rabbit IgG and horse anti-mouse IgG secondary antibodies were applied for IL-3 and IL-3Rɑ respectively (1:100, BA-1000 and BA-2000, Vector Laboratories) followed by streptavidin DyLight 594 (1:600, SA-5594, Vector Laboratories).

Techniques: Flow Cytometry, Staining

Journal: Nature

Article Title: Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease

doi: 10.1038/s41586-021-03734-6

Figure Lengend Snippet: a. Representative immunofluorescence images of the cortex of control subjects (C) or Alzheimer’s disease (AD) patients stained for IL-3 and GFAR b. Measurement of IL-3 levels in cortex tissue homogenates from control and AD patients (n=15 controls, n=23 AD patients, see Extended Data Table S6 for characteristics). c. Representative immunofluorescence images of the cortex of humans with or without Alzheimer’s disease (AD) stained for IL-3Rɑ and Iba1. d. qPCR analysis of fold change in IL3Rɑ expression in the frontal cortex of control non-demented individuals and AD patients (n=28 controls, n=30 AD patients, see Extended Data Table S7 for characteristics). e. Assessment of brain IL3Rɑ expression with APOE genotype (n=30 AD patients). f. Correlation of mean IL3Rɑ expression with years (yrs) of disease duration (n=30 AD patients). Correlation of IL3Rɑ expression with formic acid (FA)-soluble Aβ40 (g) and Aβ42 (h) in the frontal cortex of AD patients (n=23 AD patients). *p<0.05, **p<0.01, ***p<0.001. Open circles represent control subjects and red circles represent AD patients. Error bars indicate mean ± SEM.

Article Snippet: Biotinylated goat anti-rabbit IgG and horse anti-mouse IgG secondary antibodies were applied for IL-3 and IL-3Rɑ respectively (1:100, BA-1000 and BA-2000, Vector Laboratories) followed by streptavidin DyLight 594 (1:600, SA-5594, Vector Laboratories).

Techniques: Immunofluorescence, Staining, Expressing

Journal: Nature

Article Title: Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease

doi: 10.1038/s41586-021-03734-6

Figure Lengend Snippet: Missense mutations detected in CRISPR Cas9 edited mice. Missense mutations do not influence EGFP signal in Il3 targeted mice but result in quenching of EGFP signaling in Il3rɑ targeted mice. IL-3Rɑ functionality is not impacted ( ).

Article Snippet: Biotinylated goat anti-rabbit IgG and horse anti-mouse IgG secondary antibodies were applied for IL-3 and IL-3Rɑ respectively (1:100, BA-1000 and BA-2000, Vector Laboratories) followed by streptavidin DyLight 594 (1:600, SA-5594, Vector Laboratories).

Techniques: CRISPR, Mutagenesis

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: Sections were prepared from skin samples of Nc/Nga mice treated with or without vaseline ointment containing either vehicle, PQA-18 or FK506, and stained by anti-PGP9.5 antibody (green) (A) or the anti-IL-31Rα antibody (green) (B) and Hoechst33342 (blue; Nuclei). Scale bar: 100 μm. The representative images are shown on the left, and quantitative data of the number of nerve fiber are shown on the right. Data are pooled from three independent experiments with nine mice per group and shown as mean and SD. * p < 0.05, as compared with lesioned and vehicle (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Staining

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: DRG cells were treated with or without rIL-31 in the absence or presence of PQA-18, and the cell morphology were examined by immunocytochemistry with anti-βIII-tubulin (green) and IL-31Rα (red) antibodies. Scale bar: 300 μm. The representative micrographs are shown (upper), and the quantitative data of the neurite outgrowth are shown (lower). Data are pooled from three independent experiments and shown as mean and SD. ** p < 0.01 as compared with control; ## p < 0.01 as compared with the rIL-31-treated group (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Immunocytochemistry

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: The expression of IL-31Rα was determined by immunoblotting with anti-IL-31Rα antibody and anti-β-actin antibody (loading control) (A). The expression of IL-31Rα was analyzed by immunocytochemistry with anti-IL-31Rα antibody (green) or anti-PGP9.5 antibody (green) and Hoechst33342 (blue; Nuclei) (B). The representative micrographs are shown. Scale bar, 100 μm. Phosphorylated STAT3 at Tyr705 was analyzed by immunoblotting. The cells were treated with rIL-31 (C) or anti-IL-31Rα antibody (D) at indicated concentrations, and the cell lysates were examined by immunoblotting with indicated antibodies. The representative images are shown (upper) and the quantitative data of the ratios of phosphorylated STAT3 versus STAT3 are shown (lower). Data are pooled from three independent experiments and shown as mean and SD. ** p < 0.01, as compared with control (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Expressing, Western Blot, Immunocytochemistry

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: Phosphorylated STAT3 at Tyr705, JAK2 at Tyr1008, and PAK2 at Ser141 were analyzed by immunoblotting. Neuro2A cells were treated with anti-IL-31Rα antibody in the absence or presence of PQA-18 at indicated concentrations, and the cell lysates were examined by immunoblotting with indicated antibodies. The representative images are shown on the left, and the quantitative data of the ratios of phosphorylated STAT3 versus STAT3, phosphorylated JAK2 versus JAK2, and phosphorylated PAK2 versus PAK2 are shown on the right. Data are pooled from three independent experiments and shown as mean and SD. ** p < 0.01, * p < 0.05 as compared with control; ## p < 0.01, # p < 0.05 as compared with anti-IL-31Rα antibody-treated group (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: Neuro2A cells were treated with anti-IL-31Rα antibody in the absence or presence of PQA-18 at indicated concentrations, and the cell morphology was examined by microscopy. Neurites were defined as a process with lengths equivalent to one diameter of a cell body. The percentage of neurite-bearing cells was calculated from the total number of counted cells. The representative micrographs are shown (A), and the quantitative data of the neurite outgrowth are shown (B). Scale bar: 20 μm. Data are pooled from three independent experiments and shown as mean and SD. ** p < 0.01 as compared with control; ## p < 0.01 as compared with anti-IL-31Rα antibody-treated group (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Microscopy

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: Neuro2A cells were treated with anti-IL-31Rα antibody in the absence or presence of FRAX597 at indicated concentrations. Phosphorylated PAK2 at Ser141 were analyzed by immunoblotting (A). The representative images are shown on the left, and the quantitative data of the ratios of phosphorylated PAK2 versus PAK2 are shown on the right. Neurites were defined as a process with lengths equivalent to one diameter of a cell body. The percentage of neurite-bearing cells was calculated from the total number of counted cells. The representative micrographs are shown (B), and the quantitative data of the neurite outgrowth are shown (C).Scale bar: 20 μm. Data are pooled from three independent experiments and shown as mean and SD. ** p < 0.01 as compared with control; ## p < 0.01, # p < 0.05 as compared with anti-IL-31Rα antibody-treated group (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: Neuro2A cells were transfected with siRNA for PAK2 or control siRNA. PAK2 expression was determined by immunoblotting (A). Representative images of Neuro2A cells after transfection with siRNA for PAK2 or control siRNA. Neurites were defined as a process with lengths equivalent to one diameter of a cell body (B). The percentage of neurite-bearing cells was calculated from the total number of counted cells. Scale bar: 20 μm. Phosphorylated STAT3 at Tyr705 was analyzed by immunoblotting (C). Neuro2A cells were treated with rIL-31 or anti-IL-31Rα antibody in the absence or presence of siRNA for PAK2, and the cell lysates were examined by immunoblotting with indicated antibodies. The representative images are shown (upper), and the quantitative data of the ratios of phosphorylated STAT3 versus STAT3 are shown (lower). Data are pooled from three independent experiments and shown as mean and SD. ** p < 0.01 as compared with control; ## p < 0.01 as compared with rIL-31-treated group; $ $ p < 0.01 as compared with anti-IL-31Rα antibody-treated group (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Transfection, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Prenylated quinolinecarboxylic acid compound-18 prevents sensory nerve fiber outgrowth through inhibition of the interleukin-31 pathway

doi: 10.1371/journal.pone.0246630

Figure Lengend Snippet: PAK2 interacting proteins as identified by LC/MS/MS (A). LC/MS/MS analysis was performed using tryptic peptides of purified PAK2 prepared from Neuro2A cells treated without or with PQA-18 in the absence or presence of anti- IL-31Rα antibody using anti-PAK2 antibody conjugated beads. The quantitative data of the peak area of the indicated proteins are shown. Immunoprecipitates were obtained from Neuro2A cells treated without or with PQA-18 in the absence or presence of anti- IL-31Rα antibody using anti-PAK2 antibody conjugated beads, and immunoblotted with anti-α-PIX antibody or anti-PAK2 antibody (B). The representative images are shown (upper), and the quantitative data of the ratios of α-PIX versus PAK2 are shown (lower). Data are pooled from three independent experiments and shown as mean and SD. ** p < 0.01, * p < 0.05 as compared with control; ## p < 0.01, # p < 0.05 as compared with anti-IL-31Rα antibody-treated group (one-way ANOVA/Tukey-Kramer post-hoc comparisons).

Article Snippet: Sections were blocked with 5% swine serum (Vector Laboratories, Burlingame, CA) and stained with rabbit anti-PGP9.5 antibody and rabbit anti-IL-31Rα antibody.

Techniques: Liquid Chromatography with Mass Spectroscopy, Purification