Ba 9400

Manufactured by Vector Laboratories

Sourced in United States, United Kingdom

The BA-9400 is a benchtop analytical centrifuge designed for a variety of laboratory applications. It features a brushless motor and digital speed control, allowing for precise speed regulation. The centrifuge can accommodate various rotor types to accommodate different sample volumes and tube sizes.

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Lab products found in correlation

20 protocols using ba 9400

Detecting Newly Divided Cells in Dentate Gyrus

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To detect BrdU-positive (newly divided) cells in the dentate gyrus. Free-floating sections were washed in tissue buffering solution (TBS; 1.3% Trizma hydrochloride, 0.19% Trizma base, 0.9% sodium chloride) and then treated with 0.6% hydrogen peroxide in TBS for 30 min. To denature DNA, sections were treated for 120 min with a solution of 50% de-ionized formamide and 2X SSC buffer, rinsed for 15 min in 2X SSC buffer, then treated with 2 M hydrochloric acid for 30 min at 37 °C, then 0.1 M boric acid in TBS (ph 8.5) for 10 min at room temperature. Sections were then treated (blocked) with a solution of 0.3% Triton-X and 3% goat serum in TBS (TBS-X plus) for 30 min, and then incubated in primary antibody against BrdU made in rat (AbD Serotec, Raleigh, NC, USA, catalog number OB0030) at a dilution of 1:100 in TBS-X plus for 72 h at 4°C. Sections were then washed in TBS, blocked with TBS-X plus for 30 min, and then incubated in biotinylated secondary antibody against rat made in goat (Vector, Burlingame, CA, USA, catalog number BA-9400) at 1:250 in TBS-X plus for 100 min at room temperature. Sections were then treated using the ABC system (Vector, Burlingame, CA, USA, catalog number PK-6100) and stained using a diaminobenzidine kit (Sigma, St. Louis, MO, USA, catalog number D4418-505ET).

Mustroph M.L., Merritt J.R., Holloway A.L., Pinardo H., Miller D.S., Kilby C.N., Bucko P., Wyer A, & Rhodes J.S. (2014). Increased adult hippocampal neurogenesis is not necessary for wheel running to abolish conditioned place preference for cocaine in mice. The European journal of neuroscience, 41(2), 216-226.

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Hippocampal Neurogenesis Quantification via BrdU

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A subset of each group (n=5–7 per group) was used to measure adult hippocampal neurogenesis using the BrdU method as described previously [23 (link)]. In brief, a 1-in-6 series of free floating sections were washed in tissue-buffering solution (TBS; 1.3% Trizma hydrochloride, 0.19% Trizma base, 0.9% sodium chloride) and then treated with 0.6% hydrogen peroxide in TBS for 30 min. To denature DNA, sections were treated for 120 min with a solution of 50% de-ionized formamide and 2×SSC buffer, rinsed for 15 min in 2×SSC buffer, then treated with 2 M hydrochloric acid for 30 min at 37 °C, then 0.1 M boric acid in TBS (pH 8.5) for 10 min at room temperature. Sections were then blocked with a solution of 0.3% Triton-X and 3% goat serum in TBS (TBS-X plus) for 30 min, and then incubated in primary antibody against BrdU made in rat (AbD Serotec, Raleigh, NC, USA, Catalog No. OBT0030) at a dilution of 1:100 in TBS-X plus for 72 h at 4 °C. Sections were then washed in TBS, blocked with TBS-X plus for 30 min, and then incubated in biotinylated secondary antibody against rat made in goat (Vector, Burlingame, CA, USA, Catalog No. BA-9400) at 1:250 in TBS-X plus for 100 min at room temperature. Sections were then treated for 60 min using the ABC system (Vector, Burlingame, CA, USA, Catalog No. PK-6100) and stained for 5 min using a DAB kit (Sigma, St. Louis, MO, USA, Catalog No.D4418).

Merritt J, & Rhodes J.S. (2014). Mouse genetic differences in voluntary wheel running, adult hippocampal neurogenesis and learning on the multi-strain-adapted plus water maze. Behavioural brain research, 280, 62-71.

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Ly-6G Immunohistochemistry in Paraffin Sections

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5-μm paraffin-embedded tissue sections were deparaffinized in xylene and rehydrated using an ethanol gradient. Tissue sections were incubated with 3% hydrogen peroxide in PBS for 30 minutes at room temperature. Epitope retrieval was performed by treating the tissues with 10 mM sodium citrate buffer (pH 6.0) with 0.05% Tween 20 at 100° C for 10 minutes in a pressure cooker. Ly-6G staining sections were blocked with 10% normal goat serum with 1% BSA in TBS for 2 h at room temperature followed by incubation with rat monoclonal anti-Ly6g antibody (1: 500 dilution) (Abcam, Cambridge, MA, USA. ab25377) in TBS with 1% BSA at 4°C overnight. Tissue sections were treated with their respective biotinylated secondary antibodies for 45 minutes at room temperature (Vector laboratories PK-6101 and BA-9400). Color was developed using the Vectastain ABC kit (Vector Laboratories) followed by DAB reaction. Sections were then counterstained with hematoxylin, dehydrated in an ethanol and xylene. Images were acquired at 20X magnification using an Olympus microscope equipped with a D-26 color camera.

Hamed L., Emilie V., Xiao B., Canup B.S., Duke G., Denning T.L, & Didier M. (2014). Fab’-bearing siRNA TNFα-loaded nanoparticles targeted to colonic macrophages offer an effective therapy for experimental colitis. Journal of controlled release : official journal of the Controlled Release Society, 186, 41-53.

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Immunohistochemical Detection of T Cell Subsets

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Brains were removed from deeply anesthetized and exsanguinated mice, frozen in liquid nitrogen and sectioned in a cryostat. Parallel brain sections (20 μm thick) were fixed 45 min in 4% paraformaldehyde for CD4 immunostaining or 4 min in 4% phosphate buffered formaline, pH 7.4, followed by 2 min in 50%, 100%, and 50% acetone, respectively, and finally air dried for CD8a immunostaining. Sections were then rinsed in 0.5 % triton X-100 diluted in 0.05M trizma base buffered saline (TBS+T), pH 7.4, and incubated with 10% fetal calf serum (FBS) in TBS+T before incubation overnight at 4°C with purified rat-anti-mouse CD4 IgG2a (2.5 μg/mL) (553647, BD Pharmingen) or purified rat-anti-mouse CD8a IgGa (7 μg/mL) (MCA1108GT, AbDSerotec) diluted in 10% FCS in TBS+T. Parallel sections were incubated with rat IgG2a isotype control (2.5–7 μg/mL) (BD Pharmingen). After a rinse in TBS+T, endogenous peroxidase activity was blocked with 1.9% H₂O₂ in TBS, and sections were rinsed in TBS+T before and after incubation with biotinylated goat anti-rat IgG antibody (5 μg/mL) (BA9400, Vector Laboratories Inc) and HRP-conjugated streptavidin (1:300) (RPN1231V, GE Healthcare), respectively, diluted in 10% FCS in TBS+T. After a final rinse in TBS+T, the colour signal was developed in 0.05% 3.30-diaminobenzidine tetrahydrochloride and 0.003% H₂O₂ in TBS. Finally, sections were coverslipped with Depex.

Bassi M.R., Kongsgaard M., Steffensen M.A., Fenger C., Rasmussen M., Skjødt K., Finsen B., Stryhn A., Buus S., Christensen J.P, & Thomsen A.R. (2014). CD8+ T cells complement antibodies in protecting against YF virus. Journal of immunology (Baltimore, Md. : 1950), 194(3), 1141-1153.

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Microglial Phagocytosis and POMC Neurons

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Animals were sacrificed at 20–21 weeks of age by perfusion fixation (see supplementary methods). Hypothalamic 30 µm thick coronal sections were selected from each mouse covering the rostral–caudal arcuate (ARC) nucleus region. Immunohistochemical staining of the ionized calcium-binding adaptor molecule 1 (Iba1)- and proopiomelanocortin (POMC)-ir were performed to profile, respectively, the microglial morphology and the neighboring POMC neurons; immunofluorescent co-staining of Iba1 and CD68 was performed to examine microglial phagocytic capacity in the ARC region. See supplementary methods for further information on staining protocols.
Primary antibodies: rabbit anti-iba1 (Ref: 234003, Synaptic Systems), rabbit anti-POMC (Ref: H-029-30, Phoenix Pharmaceuticals), rat anti-CD68 (ab53444, Abcam).
Secondary antibodies: goat anti-rabbit IgG biotinylated (Ref: BA-1000, Vector Laboratories), goat anti-rat IgG, biotinylated (Ref: BA-9400, Vector Laboratories). Fluorescent secondary antibodies: Streptavidin, Alexa Fluor 488 (S32354, Invitrogen), goat anti-rabbit IgG, Alexa Fluor 594 (A11037, Invitrogen).

Milanova I.V., Korpel N.L., Correa-da-Silva F., Berends E., Osman S., la Fleur S.E., Fliers E., Kalsbeek A, & Yi C.X. (2022). Loss of Microglial Insulin Receptor Leads to Sex-Dependent Metabolic Disorders in Obese Mice. International Journal of Molecular Sciences, 23(6), 2933.

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Histological and Immunohistochemical Analysis of Raman Imaging Samples

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After Raman imaging, the tissues were fixed in 4% paraformaldehyde (4 °C overnight) and subsequently processed to be embedded in paraffin. Tissue sections (5 μm) were stained with hematoxylin and eosin (H&E), and immunohistochemistry staining was performed using the DISCOVERY XT biomarker platform (Ventana, Tucson, AZ, USA). Heat-induced epitope retrieval was performed using citrate buffer (pH 6.0). The primary antibodies were diluted as follows: anti-CD44 antibody (1:500; #550538; BD Biosciences, San Jose, CA, USA), anti-Ki-67 antibody (1:250, VP-RM04, Vector Laboratories Inc., Burlingame, CA, USA), anti-CCND1 antibody (1:500; Abcam, Cambridge, MA, USA), and anti-PEG antibody (1:100, PEG-B-47, ab51257, Abcam). All biotin-labeled secondary antibodies, including anti-rabbit antibody (1:300, BA-1000) and anti-rat antibody (1:300, BA-9400), were purchased from Vector Laboratories. All murine specimens were examined by a veterinary pathologist (JRW) from the Tri-Institutional Laboratory of Comparative Pathology who was blinded to the Raman images. All rat specimens were examined by a pathologist (MvdR) from the Department of Pathology at Stanford University who was blinded to the Raman images.

Harmsen S., Rogalla S., Huang R., Spaliviero M., Neuschmelting V., Hayakawa Y., Lee Y., Tailor Y., Toledo-Crow R., Kang J.W., Samii J.M., Karabeber H., Davis R.M., White J.R., van de Rijn M., Gambhir S.S., Contag C.H., Wang T.C, & Kircher M.F. (2019). Detection of Premalignant Gastrointestinal Lesions Using Surface-Enhanced Resonance Raman Scattering−Nanoparticle Endoscopy. ACS nano, 13(2), 1354-1364.

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Quantitative Immunohistochemistry Analysis

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The tissues from the imaging studies were collected and fixed in 4% paraformaldehyde, 4°C overnight and subsequently processed to be embedded in paraffin. The Discovery XT biomarker platform (Ventana) was used to stain the tissue sections (5 μm). Heat-induced epitope retrieval was performed using citrate buffer (pH 6.0). The primary antibodies were diluted as follows: anti-polyomavirus medium T antigen (PyMT) antibody (1:800, ab15085, Abcam); anti-PEG antibody (1:100, PEG-B-47, ab51257, Abcam); anti-Ki67 antibody (1:250, VP-RM04, Vector Laboratories); anti-Vimentin antibody (1:5000, V6389, Sigma-Aldrich); anti-cytokeratin 19 antibody (1:5000, 3863-1, Epitomics); anti-c-Myc (C-19) antibody (1:100, sc-788, Santa Cruz); and anti–androgen receptor (N-20) antibody (1:150, sc-816, Santa Cruz). All biotin-labeled secondary antibodies, including anti-rabbit antibody (1:300, BA-1000), anti-rat antibody (1:300, BA-9400), and anti-mouse antibody (1:300, BA-9200) were purchased from Vector Laboratories. All histological results were reviewed by two experienced mouse pathologists (J.R.W. and S.M.) from the Tri-Institutional Laboratory of Comparative Pathology who were blinded to the Raman data and used published consensus reports for lesion grading (42 (link)).

Harmsen S., Huang R., Wall M.A., Karabeber H., Samii J.M., Spaliviero M., White J.R., Monette S., O’Connor R., Pitter K.L., Sastra S.A., Saborowski M., Holland E.C., Singer S., Olive K.P., Lowe S.W., Blasberg R.G, & Kircher M.F. (2015). Surface-Enhanced Resonance Raman Scattering Nanostars for High Precision Cancer Imaging. Science translational medicine, 7(271), 271ra7.

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Evaluating Blood-Brain Barrier Permeability

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To assess changes in blood-brain barrier (BBB) permeability, sections encompassing the entire rostro-caudal axis of the brain from Baseline Cohort brains (chow, n = 8 and fructose, n = 10) were stained for immunoglobulin G (IgG; 8–12 sections per animal, section sampling fraction (ssf) = 1/12). After blocking, sections were incubated in goat anti-rat IgG (1:1000, BA9400, Vector Labs, Burlingame, CA) overnight at 4 °C, washed, incubated with the Vectastain Elite ABC kit (Vector Labs, Burlingame, CA), and visualized with diaminobenzidine (SigmaFast 3,3′diaminobenzidine tablets, Sigma Aldrich, St. Louis, MO). Increased cerebral IgG presence indicates increased blood-brain-barrier (BBB) permeability, as endothelial cell tight junctions typically prevent transport of large molecules such as IgG across the BBB (Abbott et al., 2010 (link)). A Nikon Eclipse 90i microscope (Melville, NY) fitted with MicroBrightField Stereoinvestigator Version 11 (MBF Bioscience, Williston, VT) was used to visualize IgG. To assess permeability, a research assistant blinded to treatment group captured images of each brain section using Stereoinvestigator and calculated the percentage of the brain immunoreactive for IgG using the Gray Level Index to quantify mean optical density for each brain (ImageJ, NIH).

Harrell C.S., Zainaldin C., McFarlane D., Hyer M.M., Stein D., Sayeed I, & Neigh G.N. (2018). High-fructose diet during adolescent development increases neuroinflammation and depressive-like behavior without exacerbating outcomes after stroke. Brain, behavior, and immunity, 73, 340-351.

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Immunohistochemistry Protocol for Rat Brain Tissue

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Rats were treated with either AU-FUS or standard₃-FUS and remained under 2% isoflurane anesthesia for 3 h while maintaining body temperature with a thermometric heating blanket with rectal probe (rats were injected with Lactated Ringers and Meloxicam). Rats were injected with ketamine (100 mg kg⁻¹) and xylazine (10 mg kg⁻¹) prior to transcardial perfusion. Following perfusion, the brains were post-fixed for ~18 h before preparing sections with a compresstome (50 μm thickness). Sections were blocked with 0.3% H₂O₂ in PBS for 10 min at room temperature (RT), rinsed with PBS containing 0.25% triton X-100 (PBST), and blocked with PBST containing 10% normal goat serum. The tissue was then incubated with Biotinylated anti-rat IgG (Vector labs BA-9400) diluted 1:1500 in PBST overnight at 4 °C. Sections were rinsed three times with PBST, incubated in PBST containing horseradish peroxidase avidin D (diluted 1:8000; Vector labs A-2004) for 1 hr at RT, and washed three times with PBST before adding a solution containing 0.05% diaminobenzidine, 0.01% H₂O₂, and 0.3% imidazole in PBS for 10 min at RT. The sections were immediately rinsed three times and mounted on glass slides before applying Fluoroshield with DAPI and sealing cover slips.

Ozdas M.S., Shah A.S., Johnson P.M., Patel N., Marks M., Yasar T.B., Stalder U., Bigler L., von der Behrens W., Sirsi S.R, & Yanik M.F. (2020). Non-invasive molecularly-specific millimeter-resolution manipulation of brain circuits by ultrasound-mediated aggregation and uncaging of drug carriers. Nature Communications, 11, 4929.

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Immunohistochemistry for Myelin Markers

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Established protocols were used for immunohistochemistry (Slowik et al., 2015 (link)). In brief, sections were rehydrated and, if necessary, antigens were unmasked with citrate buffer (pH 6.0) and microwave heating. After washing in PBS, sections were incubated overnight (4°C) with the primary antibodies diluted in blocking solution (serum of species in which the secondary antibody was produced). The following primary antibodies were used: anti‐PLP (1:5000, Serotec, MCA839G; Puchheim, Germany), anti‐MBP (1:500, Abcam, ab7349; Cambridge, UK), and anti‐OLIG2 (Millipore). The next day, slides were incubated with biotinylated secondary antibodies (both 1:200, Vector Labs; anti‐mouse, BA9200; anti‐rat, BA9400) for 1 h and then with peroxidase‐coupled avidin‐biotin complex (ABC kit; Vector Laboratories, Peterborough, UK) and treated with 3,3′‐diaminobenzidine as a peroxidase substrate. Stained sections were digitalized using a Nikon ECLIPSE E200 microscope (Nikon Instrument) equipped with a DS‐2Mv camera.

Lindsay S.L., McCanney G.A., Zhan J., Scheld M., Smith R.S., Goodyear C.S., Yates E.A., Kipp M., Turnbull J.E, & Barnett S.C. (2023). Low sulfated heparan sulfate mimetic differentially affects repair in immune‐mediated and toxin‐induced experimental models of demyelination. Glia, 71(7), 1683-1698.

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