Prb 278p

Manufactured by BioLegend

Sourced in United States

The PRB-278P is a precision multi-channel pipette that can accurately and precisely dispense volumes ranging from 0.5 to 10 μL. It features an ergonomic design and is made of high-quality materials for reliable and consistent performance.

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PRB-278p-100 (2 citations) Pax6 (PRB-278P, (2 citations) Rabbit anti-PAX6 (PRB-278P, (2 citations)

7 protocols using prb 278p

Antibody Staining of Neural Markers

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Anti-POGZ was prepared as described^{26 (link)}; chicken anti-GFP (Aves Laboratories GFP‐1020 1:1000); rabbit anti-Ki67 (Thermo Scientific RB-1510 1:1000); rabbit anti-Pax6 (BioLegend PRB 278P 1:500); rabbit anti-TBR2 (ab23345 1:250); rabbit anti-phospho-histone H3 (Cell Signaling Technology 9701 1:400) Alexa Fluor® 488-labeled IgG was used as secondary antibody (ab150077 1:200).

Suliman-Lavie R., Title B., Cohen Y., Hamada N., Tal M., Tal N., Monderer-Rothkoff G., Gudmundsdottir B., Gudmundsson K.O., Keller J.R., Huang G.J., Nagata K.I., Yarom Y, & Shifman S. (2020). Pogz deficiency leads to transcription dysregulation and impaired cerebellar activity underlying autism-like behavior in mice. Nature Communications, 11, 5836.

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Immunofluorescence Analysis of Mouse Eye Sections

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For frozen sections, enucleated eyeballs from P0 mice and embryos were fixed in 4% paraformaldehyde for 30 min at room temperature and 30% sucrose dehydrated over night at 4°C, then embedded in OCT, frozen, and sectioned at 10 µm. Cryosections were permeabilized in 0.5% Triton X-100/PBS for 2 min, blocked in 5% donkey serum and 5% BSA in PBS for 1 h at room temperature, then incubated with primary antibodies overnight at 4°C. After rinsing, the sections were incubated with fluorescence-labeled secondary antibodies (Alexa Fluor 488, or Alexa Fluor 546, Invitrogen, Carlsbad, CA, USA) for 2 h at room temperature and counterstained with DAPI (1:1,000, H-1200, Vector Labs, Burlingame, CA, USA). Primary antibodies used for immunofluorescence were LSS (1:300, 18693-1-AP, Proteintech, Wuhan, China), p57^KIP2 (1:200, ab75974, Abcam, Cambridge, MA, USA), Pax6 (1:200, PRB-278P, BioLegend, San Diego, CA, USA), and Prox1 (1:200, 11067-2-AP, Proteintech). The images were captured with an LSM980 confocal scanning microscope (Carl Zeiss, Thornwood, NY, USA) or a TissueFAXS microscope (TissueGnostics, Austria).

Zhao M., Mei T., Shang B., Zou B., Lian Q., Xu W., Wu K., Lai Y., Liu C., Wei L., Zhu J., Zhang K., Liu Y, & Zhao L. (2021). Defect of LSS Disrupts Lens Development in Cataractogenesis. Frontiers in Cell and Developmental Biology, 9, 788422.

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Immunofluorescence Staining of Frozen Tissue

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Frozen sections were washed in PBS with 0.2% Triton X-100 (PBST) and blocked with 5% normal donkey serum (NDS) for 1 h. The slides were then incubated with primary antibodies mixed in 1% NDS overnight at 4°C. Antibodies against TCF7L2 (1:500; 2569, Cell Signaling Technology), Ca_v3.1 (1:500; MABN464, Sigma-Aldrich, NeuroMab clone N178A/9), β-galactosidase (1:100; AB986, Merck Millipore), L1CAM (1:500; MAB5272, Merck Millipore), PAX6 (1:100; PRB-278P, BioLegend), KI-67 (1:100; AB9260, Merck Millipore), TUJ1 (1:65; MAB1637, Merck Millipore), NKX2-2 (1:50; 74.5A5, Developmental Studies Hybridoma Bank), SIX3 (1:100; 200-201-A26S, Rockland), POU4F1 (1:300; Fedtsova and Turner, 1995 (link)) were used. Sections were then incubated for 1 h with appropriate secondary antibody conjugated with Alexa Fluor 488 or 594 (1:500; A-21202, A-21207 and A-11076, Thermo Fisher Scientific). The slides were additionally stained with Hoechst 33342 (1:10,000; 62249, Thermo Fisher Scientific), washed and mounted with Vectashield Antifade Mounting Medium (H1000, Vector Laboratories).

Lipiec M.A., Bem J., Koziński K., Chakraborty C., Urban-Ciećko J., Zajkowski T., Dąbrowski M., Szewczyk Ł.M., Toval A., Ferran J.L., Nagalski A, & Wiśniewska M.B. (2020). TCF7L2 regulates postmitotic differentiation programmes and excitability patterns in the thalamus. Development (Cambridge, England), 147(16), dev190181.

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Immunofluorescence Staining Protocol for Transfected Cells

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Immunofluorescence staining was done 24 or 48 h after transfection or electroporation. Cells were rinsed once with 1x PBS and fixed with 4% PFA at room temperature for 15 min. Fixed cells or brain slices were rinsed with 1x PBS, incubated with blocking buffer (1x PBS with 0.3% Triton-X-100 and 5% donkey serum) at room temperature for 30 min, and further incubated with primary antibodies diluted in blocking buffer overnight at 4 °C. After three brief rinses in 1x PBS, slides were incubated for 1 h at room temperature with fluorophore-conjugated secondary antibodies (Donkey anti-chicken 488 (Jackson ImmunoReseach, 703-546-155), donkey anti-mouse 488 (Thermo Scientific, A21202), donkey anti-rabbit 594 (Thermo Scientific, A21207), donkey anti-rat 647 (Thermo Scientific, A48272); 1:1000 dilution for all). Slides were scanned with a Leica SP5 inverted confocal microscope. The following primary antibodies were used: anti-HA (Millipore 3F10 clone, rat, 1:1000), anti-GFP (Abcam ab13970, chick, 1:2000), anti-PUM2 (Bethyl A300-202A, rabbit, 1:1000), anti-CD24-PE (BioLegend 138504, rat, 1:1000), anti-CD133-APC (BioLegend 141208, rat, 1:1000), anti-PAX6 (Covance PRB-278P, rabbit, 1:500), anti-APP (BioLegend 802803, mouse, 1:1000).

Ruan X., Hu K, & Zhang X. (2023). PIE-seq: identifying RNA-binding protein targets by dual RNA-deaminase editing and sequencing. Nature Communications, 14, 3275.

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Immunohistochemistry of Primate Brain Tissue

Cited 2 times

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Mouse brain tissue was fixed overnight at 4°C in 4% PFA. Fixed macaque and mouse tissue were sectioned at 50 um using a Vibratome. Tissue was permeabilized and blocked in 3% BSA and 0.3% Triton X-100 in PBS for mouse and ferret tissue, and 5% Normal Donkey Serum with 1% BSA for macaque and human tissue. Primary antibodies were diluted in blocking buffer and incubated overnight 1-2 nights at 4°C. Primary antibodies included goat anti-SOX2 (1:200, Santa Cruz sc-170320 and 1:150 R+D Systems AF2018), chicken (1:200; Millipore AB15894) and rabbit (1:200; abcam ab23345) anti-TBR2 (1:500 Abcam ab183991), rabbit anti-PAX6 (1:200; Covance PRB-278P, 1:200 Biolegend 901301, mouse anti-HOPX (1:200; Santa Cruz sc-30216), rabbit (1:200; Novus NBP2-13800) and mouse (1:200; Novus H00010842-B01P) anti-PPP1R17, mouse anti-SATB2 (1:200; abcam ab51502), rabbit anti-Ki67 (1:200; abcam ab15580, and 1:200 Invitrogen 14-5698-2), rabbit anti-TBR1 (1:500, abcam ab31940), and mouse anti-PCNA (1:200; Millipore AB93501). Sections were then stained with Alexa secondary antibodies and DAPI. Ferret and human brain sections were processed as previously described (Johnson et al., 2015 (link), 2018 (link)). Imaging was performed using a Zeiss LSM 700 confocal microscope.

Girskis K.M., Stergachis A.B., DeGennaro E.M., Doan R.N., Qian X., Johnson M.B., Wang P.P., Sejourne G.M., Nagy M.A., Pollina E.A., Sousa A.M., Shin T., Kenny C.J., Scotellaro J.L., Debo B.M., Gonzalez D.M., Rento L.M., Yeh R.C., Song J.H., Beaudin M., Fan J., Kharchenko P.V., Sestan N., Greenberg M.E, & Walsh C.A. (2021). Rewiring of human neurodevelopmental gene regulatory programs by Human Accelerated Regions (HARs). Neuron, 109(20), 3239-3251.e7.

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Immunoblotting and Immunostaining Protocols for Neuronal Markers

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The following primary antibodies were used for immunoblotting: 1:1000 RAB35 (rabbit; Cell Signaling, 9690 S), 1:10,000 Actin [C4] (mouse; Merck-Millipore, MAB1501), 1:10,000 GAPDH [6C5] (mouse; Merck-Millipore, MAB374), 1:1000 contactin-2/TAG-1 (goat; R&D systems, AF4439), 1:1000 CHL1 (goat; R&D systems, AF2147), and 1:1000 N-cadherin [32/N-cadherin] (rabbit; BD, 610920). The following primary antibodies were used for immunostaining: 1:100 NeuN [A60] (mouse; Merck-Millipore, MAB377), 1:200 GFAP (rabbit, Frontier Institute, GFAP-Rb-Af800), 1:200 β-galactosidase (mouse, Promega, Z3781), 1:100 Cux1 (rabbit; Santa Cruz, sc13024), 1:100 Ctip2 (rat; Abcam, ab18465), 1:300 phosphorylated neurofilament [SMI-31] (mouse; Covance, SMI-31R), 1:500 Tbr2 (rabbit; Abcam, ab23345), 1:300 Pax6 (rabbit; BioLegend, PRB-278P), 1:200 phospho-Histone H3 (Ser10) [RR002] (rabbit; Cell Signaling, 9701), and 1:100 Nestin [Rat-401] (mouse; Invitrogen, 14-5843-82). The following secondary antibodies were used for immunostaining: goat anti-rat Alexa Fluor-488, donkey anti-mouse Alexa Fluor-488, donkey anti-rabbit Alexa Fluor-488, donkey anti-mouse Alexa Fluor-594, and donkey anti-rabbit Alexa Fluor-594 (all antibodies are from Life Technologies).

Maejima I., Hara T., Tsukamoto S., Koizumi H., Kawauchi T., Akuzawa T., Hirai R., Kobayashi H., Isobe I., Emoto K., Kosako H, & Sato K. (2023). RAB35 is required for murine hippocampal development and functions by regulating neuronal cell distribution. Communications Biology, 6, 440.

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Immunohistochemical Analysis of Mouse Brain

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Mice brains were fixed (E17) in Antigen Fix (Diapath), Brain slices were sectioned coronally (80μm) on a vibratome (Leica microsystems). Brain slices were washed with PBS (Phosphate-buffered saline pH 7.4) and stained in PBS 0.3% Triton X-100 supplemented with 5% of donkey serum. Primary antibodies were incubated overnight at 4°C, sections were then washed with PBS and incubated in secondary antibodies for 2 hours at room temperature. Antibodies used in this study were: Abba (homemade), Vimentin (Millipore, MAB3400), KI67 (Millipore, AB9260), phospho-histone H3 (Abcam, ab14955), Pax6 (biolegend, PRB-278P).

Carabalona A., Kallo H., Gonzalez M., Andriichuk L., Elomaa E., Molinari F., Fragkou C., Lappalainen P., Wessels M.W., Saarikangas J., & Rivera C. (2022). Identification of novel microcephaly-linked protein ABBA that mediates cortical progenitor cell division and corticogenesis through NEDD9-RhoA.

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