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Mouse anti parvalbumin

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Mouse anti-parvalbumin is a primary antibody that specifically binds to the calcium-binding protein parvalbumin. Parvalbumin is a widely used marker for a subpopulation of inhibitory interneurons in the central nervous system. This antibody can be used in various immunological techniques, such as immunohistochemistry and immunocytochemistry, to identify and study parvalbumin-expressing cells.

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Lab products found in correlation

Rat anti somatostatin, by Merck Group (5 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (3 mentions) Mouse anti neun, by Merck Group (3 mentions) Rabbit anti cleaved caspase 3, by Cell Signaling Technology (2 mentions) Rabbit anti gaba, by Merck Group (2 mentions) Chicken anti gfp, by Abcam (2 mentions) Rabbit anti gfap, by Agilent Technologies (2 mentions) Goat anti sox2, by Santa Cruz Biotechnology (2 mentions) Goat anti prestin, by Santa Cruz Biotechnology (2 mentions) Rabbit anti satb2 antibody, by Abcam (2 mentions) Alexa 488 conjugated donkey antibody against rabbit igg, by Jackson ImmunoResearch (2 mentions) Cy2 conjugated donkey antibody against mouse or rat igg, by Jackson ImmunoResearch (2 mentions) Bz x710, by Keyence (2 mentions) Tcs sp8, by Leica Microsystems (2 mentions) Rem 700, by Yamato Scientific (2 mentions) Alexa 488, by Thermo Fisher Scientific (2 mentions) Neuroexplorer, by MBF Biosciences (2 mentions) Neurolucida 360, by MBF Biosciences (2 mentions) Rabbit anti ankyrin g, by Santa Cruz Biotechnology (2 mentions) Vt1000s vibratome, by Leica camera (2 mentions)

29 protocols using mouse anti parvalbumin

1

Immunostaining of Brain Samples

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Immunostaining of brain sections or dissociated cells was performed as described previously26 (link),35 (link),64 (link). Primary antibodies used were mouse anti-ARID1B (Abcam, ab57461; Abnova, H00057492-M02), rabbit anti-cleaved caspase-3 (Cell Signaling Technology, #9664), mouse anti-BrdU (BD Biosciences, #555627), rabbit anti-p-histone-H3 (Cell Signaling Technology, #9701), rabbit anti-Ki67 (Cell Signaling, #9129), rabbit anti-β-catenin (Cell Signaling, #8480), chicken anti-GFP (Invitrogen, A10262), mouse anti-Parvalbumin (Sigma-Aldrich, MAB1572), rabbit anti-Cux1 (Sigma-Aldrich, SAB2105137), and rabbit anti-Tbr2 (Sigma-Aldrich, AB2283). Appropriate secondary antibodies conjugated with Alexa Fluor dyes (Invitrogen, A32731, A32727, A32723, A32732) were used to detect primary antibodies. DAPI (Sigma-Aldrich, D9542) was used to stain nuclei. No antigen retrieval was performed in this study.
Moffat J.J., Jung E.M., Ka M., Jeon B.T., Lee H, & Kim W.Y. (2021). Differential roles of ARID1B in excitatory and inhibitory neural progenitors in the developing cortex. Scientific Reports, 11, 3856.
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2

Immunostaining of Parvalbumin-Expressing Neurons

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Paraformaldehyde fixed and cryoprotected tissue was frozen and cut into 40 µm sections on a HM 400 Cryotome (Microm). Sections were blocked in PBS and 10% normal goat serum for 1 h at room temperature, and incubated in primary antibody overnight at 4°C (mouse anti-parvalbumin; 1:1000; Sigma-Aldrich). Sections were rinsed twice at room temperature for 5 min and then incubated in solution containing fluorescent secondary antibody at room temperature for 1 h (goat anti-mouse IgG Alexa Fluor 568; 2 µg/ml; Invitrogen). After rinsing twice in PBS for 5 min, sections were mounted on SuperfrostPlus slides (Fisher Scientific) and coverslipped using Vectashield Mounting Medium (Vector Laboratories). Images were captured on a LSM 510 Confocal Laser Scanning Microscope (Zeiss).
Brill J., Mattis J., Deisseroth K, & Huguenard J.R. (2016). LSPS/Optogenetics to Improve Synaptic Connectivity Mapping: Unmasking the Role of Basket Cell-Mediated Feedforward Inhibition. eNeuro, 3(4), ENEURO.0142-15.2016.
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3

Multimodal Neuroimaging Antibody Panel

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We used the following antibodies: rabbit anti-GFP (1:2000, Molecular Probe), rabbit anti-Fos (1:4000, Oncogene), chicken anti-GFP (1:2000, Abcam), rabbit anti-GABA (1:2000, Sigma), mouse anti-parvalbumin (1:2000, Sigma) and rabbit anti-Neuropeptide Y (1:2000, gift from J. Allen).
Cevikbas F., Braz J.M., Wang X., Solorzano C., Sulk M., Buhl T., Steinhoff M, & Basbaum A.I. (2017). Synergistic antipruritic effects of GABA-A and GABA-B agonists in a mouse model of atopic dermatitis. The Journal of allergy and clinical immunology, 140(2), 454-464.e2.
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4

Immunohistochemical Analysis of Neural Markers

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We used the following antibodies: rabbit anti-GFP (1:2000; Molecular Probes), chicken anti-GFP (1:2000; Abcam), mouse anti-parvalbumin (1:2000; Sigma-Aldrich), rabbit anti-GFAP (1:20 000; Dako), rabbit anti-Iba1 (1:1000; Wako), rat anti-somatostatin (1:5000; Millipore), mouse anti-GABA (1:4000; Sigma-Aldrich), and mouse anti-NeuN (1:2000; Chemicon).
Immunohistochemistry was performed as described previously (Braz et al., 2012 (link)). Sections were viewed with a Nikon Eclipse fluorescence microscope, and images were collected with a Zeiss camera (Axiocam). High-resolution confocal images taken on a Zeiss confocal confirmed that the immunoreactivity was cytoplasmic (0.8 µm optical sections). Brightness and contrast were adjusted using ImageJ (Version 1.49b).
Juarez-Salinas D.L., Braz J.M., Etlin A., Gee S., Sohal V, & Basbaum A.I. (2019). GABAergic cell transplants in the anterior cingulate cortex reduce neuropathic pain aversiveness. Brain, 142(9), 2655-2669.
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5

Immunohistochemistry of Inner Ear Markers

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Antigen retrieval was performed on all paraffin sections prior to staining by incubating in 10 mM Sodium Citrate Buffer (pH 6, 0.05% Tween 20) for 20 minutes at 98°C. Sections were incubated in primary antibodies overnight at 4°C, secondary antibodies for 2 hours at room temperature and DAPI nucleic acid stain (1∶24,000) for 8 minutes. The primary antibodies used were as follows: rabbit anti-MYO6 (Proteus Biosciences), rabbit anti-calretinin (Millipore), mouse anti-parvalbumin (Sigma), goat anti-prestin (Santa Cruz), goat anti-SOX2 (Santa Cruz) anti-P27KIP1 (Lab Vision), anti-PROX1 (Chemicon), anti-Na-K-ATPase α1 (Millipore), and anti-β-galactosidase (Abcam).
Savoy-Burke G., Gilels F.A., Pan W., Pratt D., Que J., Gan L., White P.M, & Kiernan A.E. (2014). Activated Notch Causes Deafness by Promoting a Supporting Cell Phenotype in Developing Auditory Hair Cells. PLoS ONE, 9(9), e108160.
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6

Histological Analysis of SIK1-MT Mice

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Histological analysis was followed by previous procedures (Mori and Morimoto, 2014 (link); Mori et al., 2019 (link)). Briefly, under deep anesthesia, 2-month-old male wild-type and SIK1-MT mice were perfused transcardially with ice-cold PBS (pH 7.4), followed by 4% paraformaldehyde in PBS. Fifty-micrometer-thick coronal sections were prepared with a sliding microtome (REM-700, Yamato Kohki Industrial). The sections were washed with PBS; blocked with PBS containing 1% bovine serum albumin, 0.1% Triton-X-100, and 10% of normal donkey serum; and incubated with mouse anti-parvalbumin (1:2000, Sigma), rat anti-somatostatin (1:200, Merck Millipore), or rabbit anti-Satb2 antibody (1:200, Abcam). After overnight incubation with primary antibodies, the brain sections were washed with PBS containing 0.1% Triton-X-100 and incubated with Alexa 488-conjugated donkey antibody against rabbit IgG or Cy2-conjugated donkey antibody against mouse or rat IgG (Jackson Immunoresearch), respectively, for 2–3 h at room temperature. After further washing with PBS, brain sections were mounted on a sliding glass, counterstained with DAPI, and coverslipped. Fluorescence images were taken with an all-in-one fluorescent microscope (BZ-X710, Keyence) and a confocal laser-scanning microscope (TCS SP8; Leica Microsystems).
Badawi M., Mori T., Kurihara T., Yoshizawa T., Nohara K., Kouyama-Suzuki E., Yanagawa T., Shirai Y, & Tabuchi K. (2021). Risperidone Mitigates Enhanced Excitatory Neuronal Function and Repetitive Behavior Caused by an ASD-Associated Mutation of SIK1. Frontiers in Molecular Neuroscience, 14, 706494.
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7

Comprehensive Immunolabeling for Neuroscience

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Primary antibodies used included the following: Guinea pig anti-VGLUT3 (1:10,000, Millipore), sheep anti-tyrosine hydroxylase (1:500, Millipore), rabbit anti-melanopsin (1:10,000, ATS), mouse anti-Brn3a (1:500, Millipore), goat anti-choline acetyltransferase (1:500, Millipore), mouse anti-NeuN (1:500, Millipore), mouse anti-GFAP (1:500, Sigma), rabbit anti-FoxP2 (1:4000, Abcam), rabbit anti-Pax2 (1:200, Zymed), rabbit anti-cleaved caspase 3 (1:100, Cell Signaling Technologies), mouse anti-parvalbumin (1:500, Sigma), and mouse anti-GAPDH (1:500, Abcam). Mouse monoclonal antibodies against γ-Pcdh proteins used for western blots (1:500–1:1000) were generated by NeuroMab in collaboration with the Weiner laboratory [55 (link)] and obtained from Antibodies, Inc.: N159/5 (detecting an epitope in constant exon 1 or 2 and thus all 22 γ-Pcdh isoforms); N144/32 (detecting all γA subfamily isoforms); N148/30 (specific for γB2); N174B/27 (specific for γC3). A rabbit polyclonal antibody raised at Affinity BioReagents against the peptide sequence VAGEVNQRHFRVDLD (within EC1) from murine γC4 was also used for western blotting (1:1000). Secondary antibodies were conjugated with Alexa-488, -568, or -647 (1:500, Invitrogen) or HRP (1:1000–1:5000, Jackson Immunoresearch).
Garrett A.M., Bosch P.J., Steffen D.M., Fuller L.C., Marcucci C.G., Koch A.A., Bais P., Weiner J.A, & Burgess R.W. (2019). CRISPR/Cas9 interrogation of the mouse Pcdhg gene cluster reveals a crucial isoform-specific role for Pcdhgc4. PLoS Genetics, 15(12), e1008554.
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8

Immunostaining of Neural Markers

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For immunostaining we followed protocols published in Ferran et al. (2015 (link)) and Garcia-Calero and Scharff (2013 (link)). The primary antibodies used in this study were: mouse anti-RC2 (1:10; Developmental Studies Hybridoma Bank, Iowa City, IA), mouse anti-TH (1:1000; Novus Biologicals), mouse anti-parvalbumin (1:2000; Sigma-Aldrich), rabbit anti-Calretinin (1:1500, SWANT). Some sections were counterstained with neutral red.
Garcia-Calero E., Martínez-de-la-Torre M, & Puelles L. (2020). A radial histogenetic model of the mouse pallial amygdala. Brain Structure & Function, 225(7), 1921-1956.
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9

Quantifying GFP+ Cells in the Organ of Corti

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To quantify GFP+ cells in the organ of Corti, hair cells were visualized using mouse anti-parvalbumin (1:1,000 overnight at 4° C, Sigma cat. #P3088) and goat anti-mouse conjugated to Alexa 568 fluorophore (1:1,000 for 1.5 hours at room temperature, ThermoFisher). Imaging was performed on a Nikon C2 confocal microscope under a 20X objective. NIS Elements Acquisition image acquisition software was used to capture z stack images of samples. Camera and laser settings were uniform for control groups, and experimental groups were imaged at the same or lower laser power and gain than control groups. All images were analyzed using ImageJ software. Images were coded and a blinded investigator counted GFP positive cells. A grid was overlaid on each image, with each square in the grid pattern demarcating 1,000 μm2. Each region counted was at least 10,000 μm2 in area and was composed of at least 10 contiguous 1,000 μm2 squares that bounded at least one parvalbumin positive hair cell. Counts were then averaged to create the total average count for the sample. Characteristics quantified included total number of GFP positive cells per 10,000 μm2 and total number of GFP positive HCs per 10,000 μm2.
Casey G., Askew C., Brimble M.A., Samulski R.J., Davidoff A.M., Li C, & Walters B.J. (2020). Self-complementarity in adeno-associated virus enhances transduction and gene expression in mouse cochlear tissues. PLoS ONE, 15(11), e0242599.
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10

Immunostaining and Morphological Analysis of Chandelier Neurons

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After imaging, slices were embedded in 4% agarose and sub-sectioned to 70 microns thick slabs using a vibratome (VT1000S vibratome, Leica). Sub-sectioned slices were suspended in the same blocking buffer as described above with primary antibodies for parvalbumin (mouse anti-parvalbumin, 1:2000, Sigma-Aldrich) and Ankyrin-G (rabbit anti-Ankyrin G, 1:500, Santa Cruz) and incubated at room temperature for two days. The blocking buffer was removed after 2 days and slices were washed with 0.1M PBS, 3 times for 15 minutes. Then, they were resuspended with blocking buffer containing secondary antibodies conjugated with fluorophores: goat anti-mouse, Alexa 405 (1:1000, Life Technologies) and goat anti-rabbit, Alexa Fluor 647(1:1000, Invitrogen) for 1 day at room temperature. On the following day, slices were washed with 0.1 M PBS, 3X for 15 min and mounted on glass slides with DABCO. Synaptic contacts were imaged with 63X and 100X oil immersion objectives using the imaging set-up described above. Morphological reconstruction: Biocytin-labeled neurons imaged before sub-sectioning were manually traced using Neurolucida-360 (MBF Bioscience) and visualized in NeuroExplorer (MBF Bioscience). In recovered chandelier neurons, strings of pre-synaptic ‘cartridge’ boutons were analyzed using Fiji (ImageJ).
Perumal M.B., Latimer B., Xu L., Stratton P., Nair S, & Sah P. (2021). Microcircuit mechanisms for the generation of sharp-wave ripples in the basolateral amygdala: A role for chandelier interneurons. Cell reports, 35(6), 109106.
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