Cb 38a

Manufactured by Swant

Sourced in Switzerland

The CB-38a is a compact benchtop centrifuge designed for general laboratory use. It features a maximum speed of 6,000 RPM and can accommodate sample volumes up to 30 mL. The centrifuge is equipped with an electronic speed control and a timer function.

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Lab products found in correlation

15 protocols using cb 38a

Immunohistochemical Analysis of Enteric Neurons

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Tissue was fixed in 4% formaldehyde overnight at 4 °C, then rinsed for 3 × 10 min in PBS. For colon tissue from mGlu5-KO mice, the mucosa, submucosal plexus, and circular muscle were carefully peeled off, leaving the LMMP. All tissue was permeabilized in 10% CASBLOCK + 0.1% Triton for 30 min and incubated in primary antisera overnight at 4 °C. Tissue was then rinsed in PBS 3 × 10 min, incubated in secondary antisera for 2 h at room temperature, then rinsed again 3× 10 min in PBS prior to being mounted on glass slides in Dako fluorescence mounting medium (Dako, Carpentaria, CA, USA) and secured with a glass coverslip.
Primary antisera used included Human anti-HuC/D (Hu; 1:5000, gift from Dr V. Lennon), Sheep anti-nNOS (1:2000, gift from P. Emson), Rabbit anti-Calbindin (1:1600, Swant #CB-38a), Goat anti-Calretinin (1:1000, Swant #CG1), and Rabbit anti-S100β (1:1000, Dako #Z0311). Secondary antisera included donkey anti-Human Alexa Fluor^® 647 (1:500, Jackson #709-605-149), donkey anti-Sheep Alexa Fluor^® 488 (1:400, Molecular Probes #A11015) and Alexa Fluor^® 647 (1:500, Molecular Probes #A21448), and donkey anti-Rabbit Alexa Fluor^® 594 (1:400, Molecular Probes #A21207) and Alexa Fluor^® 647 (1:400, Molecular Probes #A31573).

Leembruggen A.J., Lu Y., Wang H., Uzungil V., Renoir T., Hannan A.J., Stamp L.A., Hao M.M, & Bornstein J.C. (2023). Group I Metabotropic Glutamate Receptors Modulate Motility and Enteric Neural Activity in the Mouse Colon. Biomolecules, 13(1), 139.

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Calbindin D-28K Antibody Detection

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This rabbit polyclonal anti-Calbindin D-28K antibody (Swant, Cat# CB38a, RRID: AB_10000340) recognizes a 27–28 kDa band in immunoblot on brain homogenate of various species including zebrafish, chicken, mouse, rat, guinea pig, and rabbit (manufacturer’s data sheet). This antibody labels horizontal cells across species and subsets of other retinal cell types (amacrine, bipolar, and ganglion cells) in mouse (Haverkamp & Wassle, 2000 (link)).

Jiang D., Burger C.A., Casasent A., Albrecht N.E., Li F, & Samuel M.A. (2019). Spatiotemporal gene expression patterns reveal molecular relatedness between retinal laminae. The Journal of comparative neurology, 528(5), 729-755.

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Immunohistochemical Analysis of Mouse Brains

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Mice used for histology were transcardially perfused with 4% paraformaldehyde in PBS and the brains extracted. Fixed brains were kept in 4% paraformaldehyde at 4°C for 24 hr and in 30% sucrose for 24–48 hr. Coronal sections were cut at 50 µm. Free floating sections were washed in TBS, permeabilized and blocked in TBS containing 10% horse serum and 0.2% Triton X-100, and incubated overnight with primary antibodies (mouse anti-reelin, 1:2000, MAB5364, Merck Millipore; rabbit anti-calbindin, 1:5000, CB-38a, Swant). Slices were washed in TBS, incubated for 2 hr with secondary antibodies (goat anti-mouse Alexa Fluor 488, 1:500, A11029, Thermo Fisher; goat anti-rabbit Alexa Fluor 488, 1:500, A11008, Thermo Fisher; goat anti-rabbit Alexa Fluor 647, 1:500, A32733, Thermo Fisher). Slices were mounted on slides (Vectashield medium with DAPI), imaged using a confocal microscope (Leica SP8) and subsequently analyzed with ImageJ.

Bitzenhofer S.H., Westeinde E.A., Zhang H.X, & Isaacson J.S. (2022). Rapid odor processing by layer 2 subcircuits in lateral entorhinal cortex. eLife, 11, e75065.

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Cerebellar Size Analysis via Histology

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Gross anatomical analyses and HE staining were performed as previously described (Akizu et al., 2013 (link)). The resulting slices were scanned with NanoZoomer-XR (Hamamatsu Photonics, Japan). We evaluated the cerebellum size difference by measuring the vermis area from mid-sagittal sections (Pogoriler et al., 2006 (link)). We identified the midline by the absence of fastigial nucleus and cerebellar peduncles. A single midline section was used to calculate the area with ImageJ. For quantification of the surface occupied by the clusters and their distribution, two mice and one slice per region were quantified per genotype. For IHC, the cerebellum was embedded in OCT. 12μm-thick slices were generated with cryostat. The following antibodies were used for IHC: NeuN (1:200, MAB377; Millipore), Calbindin (1:10.000; CB-38A, Swant), GFAP (1:1.000; AB5804, Millipore), laminin (1:33, L9393, Sigma) and BiP (1:50, ADI-SPA-826, Enzo). All samples were mounted with ProLong Gold Antifade Mountant with DAPI (Life Tech). Images were taken with confocal Leica SP8 STED and analyzed with ImageJ.

Medina-Cano D., Ucuncu E., Nguyen L.S., Nicouleau M., Lipecka J., Bizot J.C., Thiel C., Foulquier F., Lefort N., Faivre-Sarrailh C., Colleaux L., Guerrera I.C, & Cantagrel V. (2018). High N-glycan multiplicity is critical for neuronal adhesion and sensitizes the developing cerebellum to N-glycosylation defect. eLife, 7, e38309.

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Immunohistochemical Localization of Calcium-Binding Protein in Harbor Porpoise Brain

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To reveal neurons containing the calcium-binding protein CB, we used the CB38a anti-CB rabbit polyclonal antibody from Swant (CB38a; RRID AB_10000340) at a dilution of 1:10,000. The pattern of staining within the neurons of the basal forebrain, diencephalon, and pons of the harbor porpoise was similar to that seen in other mammals (Gritti et al., 2003 (link); Bunce et al., 2013 (link); Bhagwandin et al., 2013 (link)).

Dell L.A., Patzke N., Spocter M.A., Siegel J.M, & Manger P.R. (2016). Organization of the Sleep-Related Neural Systems in the Brain of the Harbour Porpoise (Phocoena phocoena). The Journal of comparative neurology, 524(10), 1999-2017.

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Immunolabeling of Synaptic Proteins

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BDNF was detected with the mouse monoclonal BDNF antibody Mab#9 (anti-BDNF; Kolbeck et al., 1999 (link)) directed against the mature domain of BDNF (Dieni et al., 2012 (link)). For postsynaptic labeling, a rabbit polyclonal antibody against Synaptopodin (“anti-synpo”; #163002; Synaptic Systems), and a guinea pig polyclonal antibody against activity-related cytoskeletal protein (“anti-Arc”; #156005; Synaptic Systems) were used. For labeling of neuropeptides within the MF projection, rabbit polyclonal antibodies against Met-enkephalin (“anti–Met-enk”; Millipore) and Cholecystokinin (anti-CCK; #P06307; Millipore) were applied. Microglia were detected with a rabbit polyclonal antibody against ionized calcium-binding adaptor molecule 1 (“anti-Iba1”; #019-19741; Wako Chemicals). Reelin was detected with a mouse monoclonal Reelin (clone G10) antibody (“anti-Reelin”; #MAB5364; Millipore), while for Calbindin-D28K (CB) labeling, a polyclonal rabbit antibody (“anti-CB”; #CB-38a; Swant) was used. 4^′6-Diamidine-2-phenylindol (DAPI, 1:1000, 1 mg/mL solution, ThermoScientific) was used for nuclear counterstaining.

Dieni S., Nestel S., Sibbe M., Frotscher M, & Hellwig S. (2015). Distinct synaptic and neurochemical changes to the granule cell-CA3 projection in Bassoon mutant mice. Frontiers in Synaptic Neuroscience, 7, 18.

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Immunohistochemistry with DAB detection

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For immunohistochemistry using 3,3′-Diaminobenzidine (DAB) detection, slides were incubated in 1% hydrogen peroxide for 30 min before blocking. Sections were blocked in 10% normal goat serum, 0.3% Triton X-100, and PBS for 1 hour at room temperature. Primary antibodies: Rabbit anti-cleaved Casp3 (1:600; Cell Signaling Technology, 9661) and rabbit anti-calbindin (1:10,000; Swant, CB-38a) were incubated O/N at 4°C in blocking solution. After washing in PBS, tissue was incubated with a secondary antibody [goat biotinylated anti-rabbit (Vector Laboratories, BA-1000)] 1:500 dilution in PBS for 2 hours at room temperature. Sections were washed in PBS and incubated for 1 hour in ABC solution (Vector Laboratories, PK-6100). Color reaction was performed using a DAB kit (Vector Laboratories, SK-4100). Sections were rinsed in PBS and dehydrated in increasing gradient of ethanol solutions (2 to 3 min each) and xylenes (3 min). After dehydration, sections were mounted with Cytoseal (Thermo Fisher Scientific, 8310-4). Sections were imaged on a Zeiss Axioskop microscope (451485 El-einsatz) using a SpotFlex camera driven by Spot software (Spot Imaging Solutions version 4.6.1.41).

Fertuzinhos S., Legué E., Li D, & Liem KF J.r. (2022). A dominant tubulin mutation causes cerebellar neurodegeneration in a genetic model of tubulinopathy. Science Advances, 8(7), eabf7262.

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Immunohistochemical Analysis of Mouse Brain

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Mice were anaesthetized with pentobarbital and perfused transcardially with 4% paraformaldehyde. The brain specimens were carefully dissected out, post‐fixed for 1 h in 4% paraformaldehyde, cryoprotected, embedded in 12% gelatin, rapidly frozen, and sectioned at 40 μm using a freezing microtome or stored at −80°C until use. Frozen sections were processed free floating using the ABC method (ABC, Vector Laboratories) or single‐, double‐, and triple‐labeling immunofluorescence (de Waard et al., 2010; Vermeij, Dollé, et al., 2016). Primary antibodies (supplier; catalogue number, RRID number; dilution) used in this study were as follows: rabbit anti‐GFAP (DAKO; Z0334, AB_10013382; 1:8000); mouse anti‐NeuN (Millipore; MAB377, AB_2298772; 1:1,000); rabbit anti‐P53 (Leica; NCL‐p53‐CMP5, AB_2744683; 1:1000); rabbit anti‐Calbindin (Swant; CB‐38a, AB_2314070; 1:20,000); mouse anti‐Calbindin (Sigma Aldrich; C9848, AB_2313712; 1:20,000); and goat anti‐FOXP2 (Santa Cruz Biotechnology; sc21069, AB_2107124; 1:1000). Alexa488‐, Cy3‐, and Cy5‐conjugated secondary antibodies raised in donkey (Jackson ImmunoResearch) diluted at 1:200 were used for confocal immunofluorescence. Sections were analyzed and photographed using an Olympus BX40 microscope. Immunofluorescence sections were analyzed using a Zeiss LSM700 confocal microscope. Mean intensities were quantified using Fiji.

Birkisdóttir M.B., Jaarsma D., Brandt R.M., Barnhoorn S., van Vliet N., Imholz S., van Oostrom C.T., Nagarajah B., Portilla Fernández E., Roks A.J., Elgersma Y., van Steeg H., Ferreira J.A., Pennings J.L., Hoeijmakers J.H., Vermeij W.P, & Dollé M.E. (2021). Unlike dietary restriction, rapamycin fails to extend lifespan and reduce transcription stress in progeroid DNA repair‐deficient mice. Aging Cell, 20(2), e13302.

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Immunostaining for Calbindin-Expressing Neurons

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To reveal neurons containing the calcium-binding protein calbindin, we used the CB38a anti-calbindin rabbit polyclonal antibody from Swant (CB38a, Swant; RRID AB_10000340) at a dilution of 1:10000. The pattern of immunoreactivity throughout the gibbon brain was similar to that seen in other mammals (e.g. Bhagwandin et al., 2013 (link); Bunce, Zikopoulos, Feinberg, & Barbas, 2013 (link); Gritti et al., 2003 (link)).

Swiegers J., Bhagwandin A., Sherwood C.C., Bertelsen M.F., Maseko B.C., Hemingway J., Rockland K.S., Molnár Z, & Manger P.R. (2018). The distribution, number and certain neurochemical identities of infracortical white matter neurons in a lar gibbon (Hylobates lar) brain.. The Journal of comparative neurology, 527(10), 1633-1653.

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Immunostaining Protocols for Neuronal Markers

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The following antibodies and dilutions were used: rabbit anti-βIV-spectrin (1:1000, lab-generated^{16 (link)}), rabbit anti-total ankyrin-G (1:1000, lab-generated^{63 (link)}), goat anti-total ankyrin-G (1:1000, lab-generated ^{16 (link)}), rabbit anti-KCNQ2N1 (1:500 ^{64 (link)}), rabbit anti-NaV (1:500, Sigma S6936), rabbit anti-neurofascin FNIII (1:500, lab generated ^{65 (link)}), mouse anti-NeuN (1:1000, Sigma MAB377), mouse anti-caspr (1:1000, Neuromab 75-001), guinea pig anti-vGAT (1:1000, Synaptic Systems 131004), mouse anti-GABA_A receptor β2-3 (1:1000, Sigma, MAB341), mouse anti-parvalbumin (1:1000, Sigma P3088), rabbit anti-calbindin (1:1000, Swant CB-38a). Antibodies against KCNQ2 were used to label heteromeric KCNQ2/3 channels at the AIS ^{64 (link)}. Fluorescently conjugated secondary antibodies Alexa Fluor 488, 568, or 647 (1:250, Life Technologies) and Alexa Fluor 594-Streptavidin (1:1000, Jackson ImmunoResearch 016-580-084). The following reagents were used: FBS, Poly-D-lysine, Laminin, Paraformaldehyde, DNase, Urea, and N-ethylmaleimide were from Sigma-Aldrich. B27 supplement, GlutaMAX, Penicillin-Streptomycin, Neurobasal-A, Hank’s Balanced Salt Solution, Trypsin, Hepes, Lipofectamine 2000 and Prolong Gold Antifade Reagent were from Life Technologies. Bovine serum albumin was from Gemini Bioproducts. Tween 20 was from Calbiochem.

Nelson A., Caballero-Florán R., Díaz J.R., Hull J., Yuan Y., Li J., Chen K., Walder K., Lopez-Santiago L., Bennett V., McInnis M., Isom L., Wang C., Zhang M., Jones K, & Jenkins P. (2018). Ankyrin-G Regulates Forebrain Connectivity and Network Synchronization via Interaction with GABARAP. Molecular psychiatry, 25(11), 2800-2817.

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