anti-bai1 antibody  (Absolute Biotech Inc)

Journal: bioRxiv

Article Title: Uncovering the electrical synapse proteome in retinal neurons via in vivo proximity labeling

doi: 10.1101/2024.11.26.625481

Figure Lengend Snippet: (A) Proteins implicated in membrane trafficking SJ2BP and Syt4 colocalize with Cx36 in AII amacrine cells. (B) Several cytoskeleton associated proteins and regulators such as MAP6, DOCK7 or GPrin1 colocalize with Cx36 in AII cell dendrites. (C) The adhesion molecule BAI1 was often colocalized with Cx36 in AII cell dendrites. (D) Often components of chemical synapses such as SHANK2 and Glur2-3 were found in the periphery of gap junction plaques in AII amacrine cells. Scale: 10 µm. Magnified inset: 1 µm.

Article Snippet: The following primary antibodies were used: Cx36, 1:250 (Clone: 8F6, MAB3045, Millipore); Cx36, 1:250 (Clone: 1E5H5, 37-4600, Thermofisher); GFP, 1:250 (A10262, Thermofisher); ZO-1, 1:250 (Clone: ZO1-1A12, 33-9100, Thermofisher); ZO-2, 1:250 (71-1400, Thermofisher); CGN, 1:250 (PA5-5561, Thermofisher); SIPA1L3, 1:100 (30544-1-AP, Proteintech); EPS15L1, 1:250 (PA5-65940, Thermofisher); HIPR1, 1:250 (AB9882,Sigma); GluR2-3, 1:200 (07-598, Sigma); Gprin1, 1:100 (13771-1-AP, Proteintech); DOCK7, 1:100 (13000-1-AP, Proteintech); MAP6, 1:100 (NBP2-14220, Novus Biologicals); Synaptotagmin4, 1:100 (105 143, Synaptic Systems); SJ2BP, 1:100 (15666-1-AP, Proteintech); BAI1, 1:100 (NB110-81586, Novus Biologicals); SHANK2, 1:500 (Synaptic Systems, 162204); SCGN, 1:250 (CSB-PA020821LA01HU, ARP); SCGN, 1:500 (RD184120100, Biovendor); Vglut1, 1:500 (AB5905, Sigma); ITNS1, 1:100-250 (PA5-115432, Thermofisher).

Techniques: Membrane

Journal: Investigative Ophthalmology & Visual Science

Article Title: Myo/Nog Cells Give Rise to Myofibroblasts During Epiretinal Membrane Formation in a Mouse Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.64.2.1

Figure Lengend Snippet: Number of Cells Labeled With Antibodies to Markers of Myo/Nog Cells and Muscle in ERMs and the Retina

Article Snippet: Myo/Nog cells were identified in tissue sections by double labeling with the anti-BAI1 G8 IgM mouse mAb and the anti-noggin goat polyclonal anti-serum (AF719; R&D Systems, Minneapolis, MN, USA), as described previously., Double labeling also was carried out with the IgM BAI1 mAb and IgG mAb antibodies to α-SMA (F3777 diluted 1:250; Sigma-Aldrich, St. Louis, MO, USA), striated muscle myosin II (ab58899 diluted 1:200; Abcam, Cambridge, MA, USA), the leukocyte markers CD68 (ab201340 diluted 1:200; Abcam), CD45 (ab10558 diluted 1:200; Abcam), and CD18 (MA1819 diluted 1:100; ThermoFisher Scientific, Waltham, PA, USA), and human nucleoli (ab190710 diluted 1:100; Abcam).

Techniques: Labeling

Journal: Investigative Ophthalmology & Visual Science

Article Title: Myo/Nog Cells Give Rise to Myofibroblasts During Epiretinal Membrane Formation in a Mouse Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.64.2.1

Figure Lengend Snippet: Number of Cells Labeled With Antibodies to Markers of Myo/Nog Cells and Muscle in the Ciliary Body, Lens, and Cornea

Article Snippet: Myo/Nog cells were identified in tissue sections by double labeling with the anti-BAI1 G8 IgM mouse mAb and the anti-noggin goat polyclonal anti-serum (AF719; R&D Systems, Minneapolis, MN, USA), as described previously., Double labeling also was carried out with the IgM BAI1 mAb and IgG mAb antibodies to α-SMA (F3777 diluted 1:250; Sigma-Aldrich, St. Louis, MO, USA), striated muscle myosin II (ab58899 diluted 1:200; Abcam, Cambridge, MA, USA), the leukocyte markers CD68 (ab201340 diluted 1:200; Abcam), CD45 (ab10558 diluted 1:200; Abcam), and CD18 (MA1819 diluted 1:100; ThermoFisher Scientific, Waltham, PA, USA), and human nucleoli (ab190710 diluted 1:100; Abcam).

Techniques: Labeling

Journal: Investigative Ophthalmology & Visual Science

Article Title: Myo/Nog Cells Give Rise to Myofibroblasts During Epiretinal Membrane Formation in a Mouse Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.64.2.1

Figure Lengend Snippet: Localization of BAI1 and Noggin in ERMs and retina. Sections of mouse eyes with PVR grades 0 ( A-C ), 5 ( D-F ), and 3 ( G-I ) were double labeled with antibodies to BAI1 ( red ) and noggin ( green ). Nuclei were stained with DAPI ( blue ). The overlap of red and green appears yellow in merged images. BAI1 and noggin were co-localized in a small number of cells in the grade 0 retina A to - C . BAI1+/noggin+ cells were present throughout the ERM D to F and throughout the retina G to I . GCL = ganglion cell layer; IPL = inner plexiform layer; INL = inner nuclear layer; OPL = outer plexiform layer; ONL = outer nuclear layer. Bar = 9 µM.

Article Snippet: Myo/Nog cells were identified in tissue sections by double labeling with the anti-BAI1 G8 IgM mouse mAb and the anti-noggin goat polyclonal anti-serum (AF719; R&D Systems, Minneapolis, MN, USA), as described previously., Double labeling also was carried out with the IgM BAI1 mAb and IgG mAb antibodies to α-SMA (F3777 diluted 1:250; Sigma-Aldrich, St. Louis, MO, USA), striated muscle myosin II (ab58899 diluted 1:200; Abcam, Cambridge, MA, USA), the leukocyte markers CD68 (ab201340 diluted 1:200; Abcam), CD45 (ab10558 diluted 1:200; Abcam), and CD18 (MA1819 diluted 1:100; ThermoFisher Scientific, Waltham, PA, USA), and human nucleoli (ab190710 diluted 1:100; Abcam).

Techniques: Labeling, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Myo/Nog Cells Give Rise to Myofibroblasts During Epiretinal Membrane Formation in a Mouse Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.64.2.1

Figure Lengend Snippet: Number of BAI1+ cells per grade in a mouse model of PVR. Tissue sections of the eye were fluorescently labeled with the anti-BAI1 G8 mAb. The numbers of BAI1+ cells were counted in each section on the inner retinal surface (IRS; (grade 2), epiretinal membrane (ERM; grades 3–6), and retina (grades 0–6) ( A ), and the cornea, ciliary body, lens, and zonules of Zinn (grades 0–6) ( B ). The values are the mean ± standard deviation (SD) of the number of BAI1+ cells/section. The numbers of sections labeled with the BAI1 mAb are indicated above the SD bar. Grade 0 (injected with PBS): 9 eyes; grades 1/2: 7 eyes; grades 3/4 10 eyes; and grades 5/6: 13 eyes. * Indicates P values ≤ 0.004.

Article Snippet: Myo/Nog cells were identified in tissue sections by double labeling with the anti-BAI1 G8 IgM mouse mAb and the anti-noggin goat polyclonal anti-serum (AF719; R&D Systems, Minneapolis, MN, USA), as described previously., Double labeling also was carried out with the IgM BAI1 mAb and IgG mAb antibodies to α-SMA (F3777 diluted 1:250; Sigma-Aldrich, St. Louis, MO, USA), striated muscle myosin II (ab58899 diluted 1:200; Abcam, Cambridge, MA, USA), the leukocyte markers CD68 (ab201340 diluted 1:200; Abcam), CD45 (ab10558 diluted 1:200; Abcam), and CD18 (MA1819 diluted 1:100; ThermoFisher Scientific, Waltham, PA, USA), and human nucleoli (ab190710 diluted 1:100; Abcam).

Techniques: Labeling, Membrane, Standard Deviation, Injection

Journal: Investigative Ophthalmology & Visual Science

Article Title: Myo/Nog Cells Give Rise to Myofibroblasts During Epiretinal Membrane Formation in a Mouse Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.64.2.1

Figure Lengend Snippet: Localization of BAI1 and muscle proteins in ERMs and the retina. Sections of mouse eyes with PVR grades 5/6 were double labeled with antibodies to BAI1 ( red ) and α-SMA ( green ) ( A-H ) or striated muscle myosin ( green ) ( I-P ). Nuclei were stained with DAPI ( blue ). The overlap of red and green appears yellow in merged images ( D, H, L , P ). Regions within the boxes of the low magnification images in A , E , I , and M are shown at higher magnification in B to D , F to H , J to L , and N to P , respectively. BAI1 and α-SMA were co-localized in the ERM located in the vicinity of a retina fold ( arrow in A ). BAI1+/ α-SMA+ cells also were present throughout the layers of the retina ( E-H ). BAI1+/myosin+ cells were located within the ERM above the retinal folds ( white arrows in I , J - L ) and in the retina ( M - P ). ONL = outer nuclear layer; OPL = outer plexiform layer; INL = inner nuclear layer; IPL = inner plexiform layer; GCL = ganglion cell layer. Bar = 9 µM in A to D , F to H , J to L , and N and O , 57 µM in I and M , and 135 µM in E .

Article Snippet: Myo/Nog cells were identified in tissue sections by double labeling with the anti-BAI1 G8 IgM mouse mAb and the anti-noggin goat polyclonal anti-serum (AF719; R&D Systems, Minneapolis, MN, USA), as described previously., Double labeling also was carried out with the IgM BAI1 mAb and IgG mAb antibodies to α-SMA (F3777 diluted 1:250; Sigma-Aldrich, St. Louis, MO, USA), striated muscle myosin II (ab58899 diluted 1:200; Abcam, Cambridge, MA, USA), the leukocyte markers CD68 (ab201340 diluted 1:200; Abcam), CD45 (ab10558 diluted 1:200; Abcam), and CD18 (MA1819 diluted 1:100; ThermoFisher Scientific, Waltham, PA, USA), and human nucleoli (ab190710 diluted 1:100; Abcam).

Techniques: Labeling, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Myo/Nog Cells Give Rise to Myofibroblasts During Epiretinal Membrane Formation in a Mouse Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.64.2.1

Figure Lengend Snippet: Localization of Myo/Nog, human, and pigmented cells in PVR . Sections of mouse eyes with PVR were double labeled with antibodies to BAI1 and hNuc ( A, C-E ), hNuc, and TUNEL reagents ( B ), and BAI1 and striated muscle myosin ( F-H , R-U ), noggin ( J-M ), or α-SMA ( N-Q ). The colors of the fluorescent secondary antibodies are indicated in the annotations. Nuclei were stained with DAPI ( blue ). The overlap of red and green appears yellow in merged images ( B , L , M , P , Q , R-U ). G to - I , M , Q , S , and U are merges of red , green , and/or blue and DIC. HNuc+ cells in the vitreous of grades 1/2 PVR were not labeled with the BAI1 mAb A . Some hNuc+ cells were apoptotic B . BAI1+ cells lacked detectable levels of hNuc in the ERM C . One hNuc+ nucleus appeared to be present in a cell with two other unlabeled nuclei ( arrow in C , enlarged in D and E ). BAI1 staining was between the three nuclei ( D , E ). BAI1 and pigment were present in a cell with a normal nucleus and a dysmorphic nucleus ( F , G ). A pigmented cell on the surface of a grade 5/6 ERM was surrounded by BAI1+/myosin- cells ( arrow in H ). A low magnification image shows pigmented cells that had invaded the retina ( arrow in I ). BAI1+/noggin+ cells in the grades 5/6 ERM did not contain pigment ( K-M ). Pigmented cells were visible on the zonule ( arrow in M ). BAI1 and α-SMA were co-localized in the ERM ( red arrow in P ) and on the internal and external surface of the lens capsule ( N-Q , white arrows in P ). Cells with pigment were present on the surface of the ERM ( arrows in Q ). Pigmented cells surrounded an aggregate of BAI1+/myosin+ cells in the retina ( R , S ) and were interspersed with differentiated Myo/Nog cells in grades 5/6 ERMs ( T , U ). INL = inner nuclear layer; IPL = inner plexiform layer; GCL = ganglion cell layer; Z = zonule. Bar = 9 µM in A-C, F-I and K-N, 10 µM in G and H; 57 µM in J, and 12 µM in O-R.

Article Snippet: Myo/Nog cells were identified in tissue sections by double labeling with the anti-BAI1 G8 IgM mouse mAb and the anti-noggin goat polyclonal anti-serum (AF719; R&D Systems, Minneapolis, MN, USA), as described previously., Double labeling also was carried out with the IgM BAI1 mAb and IgG mAb antibodies to α-SMA (F3777 diluted 1:250; Sigma-Aldrich, St. Louis, MO, USA), striated muscle myosin II (ab58899 diluted 1:200; Abcam, Cambridge, MA, USA), the leukocyte markers CD68 (ab201340 diluted 1:200; Abcam), CD45 (ab10558 diluted 1:200; Abcam), and CD18 (MA1819 diluted 1:100; ThermoFisher Scientific, Waltham, PA, USA), and human nucleoli (ab190710 diluted 1:100; Abcam).

Techniques: Labeling, TUNEL Assay, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Myo/Nog Cells Give Rise to Myofibroblasts During Epiretinal Membrane Formation in a Mouse Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.64.2.1

Figure Lengend Snippet: Number of Cells Labeled With Antibodies to Leukocyte Markers in PVR

Article Snippet: Myo/Nog cells were identified in tissue sections by double labeling with the anti-BAI1 G8 IgM mouse mAb and the anti-noggin goat polyclonal anti-serum (AF719; R&D Systems, Minneapolis, MN, USA), as described previously., Double labeling also was carried out with the IgM BAI1 mAb and IgG mAb antibodies to α-SMA (F3777 diluted 1:250; Sigma-Aldrich, St. Louis, MO, USA), striated muscle myosin II (ab58899 diluted 1:200; Abcam, Cambridge, MA, USA), the leukocyte markers CD68 (ab201340 diluted 1:200; Abcam), CD45 (ab10558 diluted 1:200; Abcam), and CD18 (MA1819 diluted 1:100; ThermoFisher Scientific, Waltham, PA, USA), and human nucleoli (ab190710 diluted 1:100; Abcam).

Techniques: Labeling

Journal: Frontiers in Neuroscience

Article Title: Acute Response and Neuroprotective Role of Myo/Nog Cells Assessed in a Rat Model of Focal Brain Injury

doi: 10.3389/fnins.2021.780707

Figure Lengend Snippet: Identification of the NS injury tract lesion site, Myo/Nog cells and dying neurons in the rat brain. The area where the needle is inserted into parietal cortex and hippocampus is shown in the hematoxylin and eosin stained section in (A) (dashed line). The NS tract and the counting area within 1 mm of the lesion is shown in (B) . Sections were double labeled with antibodies to BAI1 (green) and TUNEL reagents (red), BAI1 (green), and Noggin (red) (D,E) , and NeuN (green) (F) and TUNEL (red) (C) . Nuclei were stained with DAPI in (B–F) . Overlap of green and red appear yellow in merged images. Unmerged images are shown as insets at the bottom of the photographs in (C–E) . Images were acquired from the uninjured posterior parietal cortex and hippocampus (A) , along the NS tract within the parietal cortex (B,D) , interface of the parietal cortex and hippocampus (C,F) , and the end of the NS tract in the hippocampus (E) . A BAI1+ Myo/Nog cell appears to have phagocytosed a TUNEL+ cell (C) . The red arrows in (C) depict separate nuclei. Myo/Nog cells co-expressed BAI1 and Noggin (D,E) . The majority of TUNEL+ cells were NeuN+ neurons (F) .

Article Snippet: Sections were double labeled with the BAI1 mAb and a goat anti-mouse polyclonal antiserum to Noggin diluted 1:200 (R&D Systems, Minneapolis, MN, United States).

Techniques: Staining, Labeling, TUNEL Assay

Journal: Frontiers in Neuroscience

Article Title: Acute Response and Neuroprotective Role of Myo/Nog Cells Assessed in a Rat Model of Focal Brain Injury

doi: 10.3389/fnins.2021.780707

Figure Lengend Snippet: Comparison of the number of Myo/Nog cells before and after NS injury and with depletion or addition of BAI1+ cells. Six tissue sections from 3 to 6 animals per treatment group were labeled with the BAI1 mAb (green in B–D ). Nuclei were stained with DAPI (blue in B–D ). BAI1+ cells were counted within a 1 mm area lateral to NS tract or in a similar area in the uninjured brain. The results are the means ± SEM 24 h after NS with injection of PBS, the BAI1 mAb and complement (BAI1ab/comp), complement alone (comp), BAI1+ cells or BAI1– cells (A) . NS injury significantly increased the number of Myo/Nog cells compared to a similar region in the uninjured brain ( ** p = 0.00001). Treatment with the BAI1 mAb and complement (abBAI1/comp) significantly reduced the number of Myo/Nog cells compared to NS with PBS ( p = 0.002), BAI1+ cells ( p = 0.00001), BAI1– cells ( p = 0.0002), or comp alone ( p = 0.04). Significantly more Myo/Nog cells were present in brains injected with Myo/Nog cells than the other groups (* p < 0.05). Myo/Nog cells were most prevalent after injection of exogenous BAI1+ cells ( D ; p = 0.00006). Unmerged images are shown as insets at the bottom of the photographs of merged images. Panels (B–D) are photographs of BAI1 and DAPI labeling in the parietal cortex of uninjured brain and NS injured brains injected with PBS or BAI1+ cells. More Myo/Nog cells were observed after injecting BAI1+ cells than in uninjured brains and those injected with PBS.

Article Snippet: Sections were double labeled with the BAI1 mAb and a goat anti-mouse polyclonal antiserum to Noggin diluted 1:200 (R&D Systems, Minneapolis, MN, United States).

Techniques: Comparison, Labeling, Staining, Injection

Journal: Frontiers in Neuroscience

Article Title: Acute Response and Neuroprotective Role of Myo/Nog Cells Assessed in a Rat Model of Focal Brain Injury

doi: 10.3389/fnins.2021.780707

Figure Lengend Snippet: Comparison of cell death before and after NS injury and with depletion or addition of BAI1+ cells. Six tissue sections from 3 to 6 animals per treatment group were stained with TUNEL reagents (red in B–D ). Nuclei were stained with DAPI (blue in B–D ). The number of TUNEL+ cells was counted within a 1 mm area lateral to NS tract or in a similar area in the uninjured brain. The results are the means ± SEM 24 h after NS with injection of PBS, the BAI1 mAb and complement (BAI1ab/comp), complement alone (comp), BAI1+ cells or BAI1– cells (A) . Significantly fewer TUNEL+ cells were present in uninjured tissue than the other treatment groups (* p = 0.00001). NS injury significantly increased the number of TUNEL+ cells in all treatment groups ( p < 0.05). Injection of BAI1+ cells isolated from the brains of other animals significantly reduced the number of TUNEL+ cells compared to all other groups ( ** p < 0.05). The numbers of TUNEL+ cells were similar in brains injected with PBS, BAI1 and complement (BAI1Ab/comp), and complement alone (Comp). Panels (B–D) are photographs of TUNEL and DAPI labeling in the parietal cortex of the uninjured brain and NS injured brains injected with PBS or BAI1+ cells. Unmerged images are shown as insets at the bottom of the photographs of merged images. TUNEL+ cells were rare in the uninjured brain (B) . Fewer TUNEL+ cells were observed after injecting BAI1+ cells (D) than in those injected with PBS (C) .

Article Snippet: Sections were double labeled with the BAI1 mAb and a goat anti-mouse polyclonal antiserum to Noggin diluted 1:200 (R&D Systems, Minneapolis, MN, United States).

Techniques: Comparison, Staining, TUNEL Assay, Injection, Isolation, Labeling

Journal: Frontiers in Neuroscience

Article Title: Acute Response and Neuroprotective Role of Myo/Nog Cells Assessed in a Rat Model of Focal Brain Injury

doi: 10.3389/fnins.2021.780707

Figure Lengend Snippet: Comparison of the number of NeuN+ neurons before and after NS injury and with depletion or addition of BAI1+ cells. Six tissue sections from 3 to 6 animals per treatment group were stained with TUNEL reagents (red in B–D ). Nuclei were stained with DAPI (blue in B–D ). NeuN positive cells were counted within a 1 mm area lateral to NS tract or in a similar area in the uninjured brain. The results are the means ± SEM 24 h after NS with injection of PBS, the BAI1 mAb and complement (BAI1ab/comp), complement alone (comp), BAI1+ cells or BAI1– cells (A) . The elevation in the number of NeuN cells in brains injected with BAI1+ compared to the uninjured brain was not statistically significant ( p = 0.06). The number of NeuN+ cells was significantly greater after injection of BAI1+ cells than in the other treatment groups (* p ≤ 0.05). Panels (B–D) are photographs of NeuN and DAPI Unmerged images are shown as insets at the bottom of the photographs of merged images.

Article Snippet: Sections were double labeled with the BAI1 mAb and a goat anti-mouse polyclonal antiserum to Noggin diluted 1:200 (R&D Systems, Minneapolis, MN, United States).

Techniques: Comparison, Staining, TUNEL Assay, Injection

Journal: Nature Communications

Article Title: Surfaceome dynamics reveal proteostasis-independent reorganization of neuronal surface proteins during development and synaptic plasticity

doi: 10.1038/s41467-020-18494-6

Figure Lengend Snippet: a cLTP paradigm b The plot shows averaged fast EPSC amplitudes for each cell (triangles) and their averages (mean ± s.e.m) for control (untreated, n = 45) and cLTP (glycine treated, n = 38) cells. Mann–Whitney test. c – f Boxplots of distributions of log2 surface abundances for replicates of cLTP ( n = 12) and control ( n = 12). Boxes indicate median and percentiles (25th and 75th). g Volcano plot of statistical significance ( y -axis) and surface abundance change ( x -axis). Horizontal line is located at an adjusted p -value of 0.05. h Proteins with significantly different surface abundances (fold-change > 1.5 and p < 0.05) after cLTP grouped into categories. Shape size indicates scaled −log10 adjusted p -value. Border color indicates fold-change directionality (green > 0, purple < 0). Edges represent string confidence > 0.7. i Representative images of analysed primary dendrites before and after cLTP induction. Bars 5 µm. j Quantification of mean fluorescent intensity of antibody signal on the primary dendrite surface using antibodies against GluA: n = 63, Bai1: n = 42, Adcy3: n = 28 (CTRL) n = 22 (cLTP) cells/group. Means + 95% CI presented. Two-way T -test (Adcy3 + Bai1) or Mann–Whitney test (GluA). * p < 0.05, ** p < 0.01, **** p < 0.0001. Source data are provided as a Source Data file.

Article Snippet: Antibodies targeting the extracellular domains of ADGRB1 (ABR-021, Alomone), AMPARs (182 411, Synaptic Systems), and AC3 (AAR-043, Alomone) were diluted 1:100 in blocking solution containing 10% normal goat serum (NGS) and incubated with the cells for 60 min.

Techniques: MANN-WHITNEY