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Rabbit anti calbindin

Manufactured by Cell Signaling Technology
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Rabbit anti-calbindin is a primary antibody that recognizes calbindin, a calcium-binding protein. Calbindin is expressed in various cell types and plays a role in calcium homeostasis. This antibody can be used to detect and analyze calbindin in biological samples through techniques such as Western blotting, immunohistochemistry, and immunocytochemistry.

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Lab products found in correlation

Rabbit anti iba1, by Fujifilm (2 mentions) Axio observer z1 microscope, by Zeiss (2 mentions) Goat anti mouse cy2, by Jackson ImmunoResearch (1 mentions) Biotinylated goat anti rabbit, by Jackson ImmunoResearch (1 mentions) Donkey anti rat cy5, by Jackson ImmunoResearch (1 mentions) Goat anti rabbit cy3, by Jackson ImmunoResearch (1 mentions) Rat anti gfap, by Thermo Fisher Scientific (1 mentions) Stable dab, by Thermo Fisher Scientific (1 mentions) Rat anti cd45, by Bio-Rad (1 mentions) Biotinylated goat anti mouse, by Jackson ImmunoResearch (1 mentions) Superfrost plus slide, by Thermo Fisher Scientific (1 mentions) Alexa fluor goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Anti mouse pkcα, by Santa Cruz Biotechnology (1 mentions) Mouse anti gapdh, by Santa Cruz Biotechnology (1 mentions) Sumitomo nerve cell culture system, by Sumitomo Bakelite (1 mentions) Mouse anti β actin, by BD (1 mentions) Rabbit anti pkcγ, by Santa Cruz Biotechnology (1 mentions) Rabbit anti pkc substrate, by Cell Signaling Technology (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Calbindin (3 citations) Anti-Calbindin (2 citations)

6 protocols using rabbit anti calbindin

1

Multimodal Analysis of Neuroinflammation

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For immunohistochemistry experiments, tissues were stained with primary antibodies directed to identify microglia (rabbit anti-Iba-1, 1:4000, Wako or mouse anti-Iba-1 kindly provided by Dr. Bruce Trapp), inflammatory monocytes (rat anti-CD45, 1:2000, Bio-Rad), cerebellar neurons (rabbit anti-calbindin, 1:1000, Cell signaling Technology), pro-inflammatory cytokine interleukin-1 beta (rabbit anti-IL1 beta, 1:200, abcam), myelin basic protein (MBP) (rabbit anti-MBP, 1:2000, Invitrogen), and astrocytes (rat anti-GFAP, 1:4000, Invitrogen). Secondary antibodies were purchased from Jackson ImmunoResearch Laboratory as follows: Cy3-goat anti rabbit, Cy5-donkey anti rat, Cy2-goat anti mouse, biotinylated-goat anti rabbit, biotinylated-goat anti mouse, and biotinylated-rabbit anti rat. Biotinylated antibodies are detected by using stable DAB (Thermo Fisher Scientific) containing diaminobenzidine and hydrogen peroxidase.
Cardona S.M., Kim S.V., Church K.A., Torres V.O., Cleary I.A., Mendiola A.S., Saville S.P., Watowich S.S., Parker-Thornburg J., Soto-Ospina A., Araque P., Ransohoff R.M, & Cardona A.E. (2018). Role of the Fractalkine Receptor in CNS Autoimmune Inflammation: New Approach Utilizing a Mouse Model Expressing the Human CX3CR1I249/M280 Variant. Frontiers in Cellular Neuroscience, 12, 365.
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2

Immunohistochemical Analysis of Olfactory Bulb

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Following intracardial perfusion with 4% PFA in PBS, brains were removed and post-fixed in 4% PFA in PBS before dissecting olfactory bulbs (OBs). Cryoprotected olfactory bulbs were sectioned into 12 μm slices using a Leica CM3050 cryostat and mounted on Fisher brand Superfrost Plus slides. Sections were permeabilized for 10 mins in a solution of 10% normal goat serum (NGS) and 0.1% Triton-X-100 in PBS, then blocked with 1% bovine albumin (BSA), 10% NGS, and 0.1% Triton-X-100 in PBS for 1 h at room temperature. Slides were incubated in primary antibody solution overnight at 4°C using: rabbit anti-parvalbumin (Swant) at 1:500, rabbit anti-tyrosine hydroxylase (Pel-Freez) at 1:1000, rabbit anti- calretinin (Swant) at 1:2000, rabbit anti-calbindin (Cell Signaling) at 1:200, and rabbit phospho-histone H3 (Cell Signaling) at 1:500. Secondary antibody (Invitrogen Alexa Fluor goat anti-rabbit, either 488 or 546) was used at 1:500 in block solution for 1 h at room temperature, then counterstained with 4′,6-diamidino-2-phenylindole, or DAPI (5 mg/mL in ddH2O), to visualize nuclei. For Calbindin staining, IHC sections underwent an additional antigen retrieval (AR) step using a 10 mM Sodium Citrate solution at 95°C for 15 mins, then placed in a blocking solution and stained as above. TUNEL staining of the olfactory bulb was done according to manufacturer's protocol (Promega).
Persson L., Witt R.M., Galligan M., Greer P.L., Eisner A., Pazyra-Murphy M.F., Datta S.R, & Segal R.A. (2014). Shh-Proteoglycan Interactions Regulate Maturation of Olfactory Glomerular Circuitry. Developmental neurobiology, 74(12), 1255-1267.
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3

Calbindin-Expressing Neurons in Mouse Brain

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Mice were euthanized at 7 weeks of age and transcardially perfused with room temperature phosphate buffered saline (PBS). The whole brain was dissected immediately and fixed in 4% paraformaldehyde (PFA) in PBS, pH 7.4, 4 °C, for 24 h followed by cryoprotection in 30% sucrose until sectioning. Brains were cryostat-sectioned parasagitally (20 μm) and sections were collected in PBS containing 0.25% Triton X-100. The floating sections were blocked for 30 min in 10% normal goat serum and then incubated overnight at 4 °C with rabbit anti-Calbindin (1:400; Cell Signaling, Danvers, MA, USA) in 0.25% Triton X-100/PBS. After washing, the sections were incubated with Alexa Fluor 594 goat anti-rabbit IgG, (1:1000, Thermo Fisher Scientific, Waltham, MA, USA). Nuclei were stained with Hoechst 33342, Trihydrochloride, Trihydrate I (1:1000, Invitrogen, Carlsbad, CA, USA). Images were taken on a Zeiss Axio Observer Z1 microscope (Zeiss, Germany).
Cawley N.X., Lyons A.T., Abebe D., Wassif C.A, & Porter F.D. (2020). Evaluation of the Potential Role of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) in Niemann–Pick Disease, Type C1. International Journal of Molecular Sciences, 21(7), 2430.
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4

Immunoblotting analysis of DGKγ, PKC, and related proteins

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Primers were purchased from Thermo Fisher Scientific. We used the following antibodies: rabbit anti-DGKγ (Adachi et al., 2005 (link)), rabbit anti-calbindin, rabbit anti-PKC substrate (Cell Signaling), rabbit anti-phospho-PKCγ T674 (Bios), rabbit anti-phospho-PKCα S657 (Abcam), rabbit anti-PKCγ, mouse anti-PKCα, mouse anti-GAPDH (Santa Cruz), mouse anti-β-actin (BD), peroxidase-conjugated AffiniPure goat anti-rabbit and anti-mouse IgG and Alexa Fluor 546 (Alexa 546)-conjugated goat anti-rabbit IgG (Jackson). We used the following cell culture reagents: Sumitomo Nerve-Cell Culture System (Sumitomo Bakelite) and 3,3′,5′-triiodo-l-thyronine (T3) sodium salt (Thermo Fisher Scientific). The plasmids, 12-o-tetradecanoylphorbol 13-acetate (TPA), GF109203X (GFX), and Gö6976 (Gö) were donated by Dr. Saito (Biosignal Research Center, Kobe University, Kobe, Japan). Scutellarin (Scu) was purchased from Namiki Shoji.
Tsumagari R., Kakizawa S., Kikunaga S., Fujihara Y., Ueda S., Yamanoue M., Saito N., Ikawa M, & Shirai Y. (2020). DGKγ Knock-Out Mice Show Impairments in Cerebellar Motor Coordination, LTD, and the Dendritic Development of Purkinje Cells through the Activation of PKCγ. eNeuro, 7(2), ENEURO.0319-19.2020.
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5

Immunofluorescent Staining of Mouse Brain

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Mice were euthanized as described above at 7 weeks of age, and transcardially perfused with phosphate buffered saline (PBS). The whole brain was dissected immediately and fixed in 4% paraformaldehyde (PFA) in PBS, pH 7.4, 4 °C, for 24 h, followed by cryoprotection in 30% sucrose until sectioning. Brains were sectioned parasagitally (20 mm) in a cryostat, and sections were collected in PBS containing 0.25% Triton X-100. The floating sections were blocked for 30 min in 10% normal goat serum in PBS, and then incubated overnight at 4 °C with rabbit anti-calbindin (1:400; Cell Signaling, Danvers, MA, USA), chicken anti-GFAP (1:1000, Novus), and rabbit anti-Iba1 (1:500, Wako Pure Chemicals, Richmond, VA, USA) in 0.25% Triton X-100/PBS. After washing, the sections were incubated with appropriate host-specific secondary antibodies coupled to either Alexa 594 and Alexa 488 (1:1000, Thermo Fisher Scientific, Waltham, MA, USA). Nuclei were stained with Hoechst 33342, trihydrochloride, and trihydrate I (1:1000, Invitrogen, Carlsbad, CA, USA). Images were taken on a Zeiss Axio Observer Z1 microscope (Zeiss, Germany).
Cawley N.X., Lyons A.T., Abebe D., Luke R., Yerger J., Telese R., Wassif C.A., Bailey-Wilson J.E, & Porter F.D. (2022). Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology. International Journal of Molecular Sciences, 23(9), 5082.
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6

Antibody Validation for Neuroscience Research

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The antibodies used were as follows: rabbit anti-actin (#926-42210, LI-COR), rabbit anti-APP (#ab32136, Abcam), mouse anti-BACE1 (3D5), rabbit anti-BACE1 (#ab108394, Abcam), rat anti-BrdU (#MA1-82088, Thermo Fisher Scientific), rabbit anti-calbindin (#2173, Cell Signaling Technology), goat anti-CHL1 (#AF2147, R&D Systems), rat anti-MBP (#ab7349, Abcam), rabbit anti-neuregulin type III (#AB5551, Millipore), chicken anti-NeuN (#ABN91, Millipore), goat anti-olfactory marker protein (OMP) (#544-1001, WAKO), mouse anti-III-tubulin (Tuj1) (a gift from L. Binder), rabbit anti-PLP (#ab28486, Abcam), goat anti-SPO (#sc21212, Santa Cruz Biotechnology), mouse anti-synaptophysin (#MAB5258, Millipore), and rat anti-Sez6 (a gift from S. Lichtenthaler).
Ou-Yang M.H., Kurz J.E., Nomura T., Popovic J., Rajapaksha T.W., Dong H., Contractor A., Chetkovich D.M., Tourtellotte W.G, & Vassar R. (2018). Axonal organization defects in the hippocampus of adult conditional BACE1 knockout mice. Science translational medicine, 10(459).
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