Anti calbindin d 28k

Manufactured by Swant

Sourced in Switzerland, Canada

Anti-Calbindin D-28k is a laboratory reagent used for the detection and quantification of Calbindin D-28k, a calcium-binding protein, in biological samples. It is commonly used in immunohistochemistry, western blotting, and other analytical techniques to identify the presence and distribution of Calbindin D-28k in various tissues and cell types.

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Lab products found in correlation

Axio observer a1, by Zeiss (3 mentions) Anti tom20, by Santa Cruz Biotechnology (2 mentions) Anti iba1, by Fujifilm (2 mentions) Anti rabbit alexa594, by Thermo Fisher Scientific (1 mentions) Streptavidin alexa 594, by Thermo Fisher Scientific (1 mentions) Anti gfp, by Thermo Fisher Scientific (1 mentions) Caspase 3, by Abcam (1 mentions) Anti rhodopsin, by Abcam (1 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (1 mentions) Anti vps26, by Abcam (1 mentions) Anti calretinin, by Swant (1 mentions) Alexa conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Anti lamp2, by Abcam (1 mentions) Anti lamp1, by Abcam (1 mentions) Anti actin, by Merck Group (1 mentions) Anti atg5, by Merck Group (1 mentions) Anti wipi2, by Abcam (1 mentions) Anti myc, by Santa Cruz Biotechnology (1 mentions) Anti lc3, by Nanotools (1 mentions) Anti calbindin d 28k, by Merck Group (1 mentions)

Spelling variants of this product

Calbindin (27 citations) Anti-calbindin (9 citations) Calbindin D-28K (4 citations)

5 protocols using anti calbindin d 28k

Immunohistochemical Analysis of Embryonic Brain

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Brains of E13.5, E16, or E18.5 embryos were processed for histology and immunohistochemistry as previously described (Lysko et al., 2011 (link)). Primary antibodies used included anti-Calbindin D-28k (rabbit; Swant, 1:1000), Caspase-3 (rabbit; Abcam, 1:500), Ki67 (rabbit; Neomarkers, 1:300), anti-Tom-20 (rabbit; Santa Cruz, 1:500), anti-GFP (chicken: Invitrogen, 1:2000). Secondary antibodies included goat anti-rabbit-biotin; Vector Laboratories followed by Streptavidin-Alexa 594; Invitrogen, or anti-rabbit-Alexa 594; Invitrogen anti-chick-Alexa 488; Invitrogen all at 1:2000. Nuclei were counterstained with DAPI.
MGE explants were fixed and immunolabeled as previously described (Lysko et al., 2011 (link); Lysko et al., 2014 (link)) using anti-Tuj1 (rabbit; Neuronal CIII b-tubulin; Covance, 1:1000), anti gamma-tubulin (mouse; 1:200 Sigma), anti-GFP (chicken Invitrogen 1:200), or anti-Ant1 (rabbit) as primary antibodies.

Lin-Hendel E.G., McManus M.J., Wallace D.C., Anderson S.A, & Golden J.A. (2016). Differential mitochondrial requirements for radially and non-radially migrating cortical neurons: Implications for mitochondrial disorders. Cell reports, 15(2), 229-237.

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Vps35 Mutant Mice Characterization

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Vps35 mutant mice have been described previously [9 (link),10 (link),14 (link)], which were backcrossed with C57BL/6 mice for more than 10 generations. Mice were maintained on a standard rodent diet. Vps35 mutations were confirmed by genotyping using PCR and Western blot analysis. All experimental procedures were approved by the Animal Subjects Committee at the Georgia Regents University according to US National Institutes of Health guidelines.
Rabbit polyclonal anti-VPS35 antibody was generated using the antigen of GST-VPS35D1 fusion protein as described previously [9 (link),10 (link),14 (link)]. Rabbit polyclonal antibodies, including anti- ß-galactosidase (Cappel), anti-VPS26 (Abcam),anti-calretinin (Swant), and anti-melanopsin (Advanced Targeting Systems) antibodies, were purchased. Mouse monoclonal antibodies, including anti-neuronal class III ß-Tubulin (Tuj1,Convance), anti-neurofilament (DSHB), anti-calbindin D28k (Swant), anti-glial fibrillary acidic protein (GFAP, Chemicon), anti-rhodopsin (Abcam) and anti-myelin basic protein (MBP, Chemicon) antibodies, were also purchased. In addition, the chicken polyclonal anti- ß-galactosidase antibody (Abcam) and goat polyclonal anti-IBA1 antibody (Abcam) were used. Secondary antibodies were purchased from Jackson Immuno Research Laboratories, Inc. Other chemicals and reagents used in this study were of analytical grade.

Liu W., Tang F.L., Erion J., Xiao H., Ye J, & Xiong W.C. (2014). Vps35 haploinsufficiency results in degenerative-like deficit in mouse retinal ganglion neurons and impairment of optic nerve injury-induced gliosis. Molecular Brain, 7, 10.

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Immunodetection of Neuronal Markers

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Cultured cells were fixed with 4% paraformaldehyde in 0.1 M PBS (pH 7.4) at room temperature (RT) for 15 min. Fixed cells were permeabilized and blocked against nonspecific protein binding (10% normal goat serum, 2% bovine serum albumin and 0.4% Triton X-100 in PBS) for 30 min at RT. The following primary antibodies were used (with specified dilution) for immunodetection: Mouse anti-neuronal nuclei (NeuN) monoclonal (1:500, Chemicon International, USA); mouse anti-tubulin, beta III isoform monoclonal (1:1000, Chemicon); anti-Calbindin D-28K (mouse monoclonal, 1:1000, Swant, Bellinzona, Switzerland and rabbit polyclonal, 1:1000, Chemicon); mouse anti-synaptophysin monoclonal (1:10,000, Chemicon). The primary antibodies were incubated overnight at 4 °C for binding, and then detected with the appropriate Alexa-conjugated secondary antibodies (Molecular Probes, Inc., 2 h incubation at RT). Actin was visualized by Alexa Fluor 594 conjugated phalloindin (1:1000, Invitrogen, USA).

Sur S., Guler M.O., Webber M.J., Pashuck E.T., Ito M., Stupp S.I, & Launey T. (2014). Synergistic regulation of cerebellar Purkinje neuron development by laminin epitopes and collagen on an artificial hybrid matrix construct. Biomaterials science, 2(6), 903-914.

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Antibody Immunodetection Assay Protocol

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The following antibodies were used for the immunoblot and immunofluorescence assays: anti-GFP (Santa Cruz Biotechnology, #sc-9996), anti-Wipi3 (Thermo Fisher Scientific, #PA5-50864), anti-Atg5 (Sigma, #A0731), anti-Lamp2 (Abcam, #ab13524), anti-RFP (MBL, #M204-3), anti-Flag (MBL, #PM020), anti-HA (Santa Cruz Biotechnology, #sc-7392), anti-Lamp1 (abcam, #ab24170), anti-VSVG (KeraFAST, #EB0010), anti-GS28 (BD, #611184), anti-GM130 (BD, #610823), anti-Tom20 (Santa Cruz Biotechnology, #sc-11415), anti-Myc (Santa Cruz Biotechnology, #sc-40), anti-Rab9 (Sigma, #R5404), anti-His (Nacalai Tesque, #04428-26), anti-LC3 (nanoTools, #0231-100), anti-p62 (MBL, #PM045), anti-Wipi2 (abcam, #ab105459), anti-actin (Millipore, #MAB1501), anti-GST (Santa Cruz Biotechnology, #sc-138), anti-Atg7 (CST, #2631), anti-GAPDH (EMD Millipore, #MAB374), anti-calbindin D-28k (Swant, #300), anti-calbindin D-28k (Sigma, #C2724), anti-GFAP (Cell Signaling Technology #12389), anti-Iba1 (Wako, #019-19741), anti-ubiquitin (CST, #3933), anti-ceruloplasmin (BD, #611488), anti-ferritin (abcam, #ab69090), anti-ferroportin (Novus, #NBP1-21502) and anti-DMT1 (Proteintech, #20507-1-AP). Enzymes used for recombinant DNA techniques were purchased from Takara Bio, TOYOBO, and New England BioLabs. All other reagents were obtained from Nacalai Tesque.

Yamaguchi H., Honda S., Torii S., Shimizu K., Katoh K., Miyake K., Miyake N., Fujikake N., Sakurai H.T., Arakawa S, & Shimizu S. (2020). Wipi3 is essential for alternative autophagy and its loss causes neurodegeneration. Nature Communications, 11, 5311.

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Immunohistochemistry of Fixed Brain Sections

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Paraformaldehyde-fixed brains were sectioned coronally (at 20μm thickness) using a sliding microtome (CM1900; Leica, Germany). For immunohistochemistry (IHC), slices were blocked with phosphate-buffered saline (PBS) containing 10% fetal bovine serum (FBS) and 0.5% Triton X-100 for 1.5 hours, and incubated with anti-GFAP (Z0334; Dako Cytomation, Denmark), anti-Iba1 (019-19741; Wako, Japan), anti-YM1 (#01404, Stemcell Technologies, Canada) and anti-Calbindin-D_28K (CB38; Swant, Switzerland) antibodies at 4°C overnight. Slices were then incubated with Alexa Fluor 488-conjugated AffiniPure Goat anti-rabbit IgG secondary antibody (111–545–003; Jackson ImmunoResearch, USA) for 1.5 hours. For Thioflavin-S staining, slices were stained with 0.015% Thioflavin-S (T1892; Sigma, USA) for 15 minutes at room temperature. After mounting, slides were imaged using a Zeiss fluorescence microscope (Axio Observer A1; Zeiss, Germany).

Tai Y.H., Lin Y.Y., Wang K.C., Chang C.L., Chen R.Y., Wu C.C, & Cheng I.H. (2018). Curcuminoid submicron particle ameliorates cognitive deficits and decreases amyloid pathology in Alzheimer’s disease mouse model. Oncotarget, 9(12), 10681-10697.

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