Recombinant Anti-Calbindin antibody [EP3478] (ab108404)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP3478] to Calbindin
- Suitable for: WB, IHC-P, ICC/IF, IHC-Fr, Flow Cyt (Intra), mIHC
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Calbindin antibody [EP3478]
See all Calbindin primary antibodies -
Description
Rabbit monoclonal [EP3478] to Calbindin -
Host species
Rabbit -
Specificity
We do not guarantee IHC-P for mouse and rat.
We do not guarantee IHC-Fr for human.
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Tested applications
Suitable for: WB, IHC-P, ICC/IF, IHC-Fr, Flow Cyt (Intra), mIHCmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Recombinant Human Calbindin protein (ab117207), fetal brain, fetal cerebellum, fetal hypothalamus and fetal pituitary lysates, SH-SY5Y, Human kidney, Mouse brain, Mouse kidney, and Rat brain lysates. IHC-P: Human cerebrum and Human brain tissue. IHC-Fr: Mouse and Rat cerebellum tissues. ICC/IF: Mouse hippocampal cells. Flow Cyt (intra): SH-SY5Y cells. mIHC: Human cerebellum tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP3478 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108404 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 30 kDa.
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IHC-P | (1) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. We do not guarantee IHC-P for mouse and rat. See IHC antigen retrieval protocols For unpurified use at 1/100 - 1/250. |
ICC/IF |
1/150.
For unpurified use at 1/100 |
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IHC-Fr | (1) |
1/500.
We do not guarantee IHC-Fr for human |
Flow Cyt (Intra) |
1/150.
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mIHC |
1/1000.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 30 kDa. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. We do not guarantee IHC-P for mouse and rat. See IHC antigen retrieval protocols For unpurified use at 1/100 - 1/250. |
ICC/IF
1/150. For unpurified use at 1/100 |
IHC-Fr
1/500. We do not guarantee IHC-Fr for human |
Flow Cyt (Intra)
1/150. |
mIHC
1/1000. |
Target
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Function
Buffers cytosolic calcium. May stimulate a membrane Ca(2+)-ATPase and a 3',5'-cyclic nucleotide phosphodiesterase. -
Sequence similarities
Belongs to the calbindin family.
Contains 5 EF-hand domains. -
Domain
This protein has four functional calcium-binding sites; potential sites II and VI have lost affinity for calcium. - Information by UniProt
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Database links
- Entrez Gene: 793 Human
- Entrez Gene: 12307 Mouse
- Entrez Gene: 83839 Rat
- Omim: 114050 Human
- SwissProt: P05937 Human
- SwissProt: P12658 Mouse
- SwissProt: P07171 Rat
- Unigene: 65425 Human
see all -
Alternative names
- avian-type antibody
- CAB27 antibody
- CALB 1 antibody
see all
Images
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Immunofluorescent analysis was performed on 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse hippocampal primary neural/glia cells labelling Calbindin with ab108404 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody (ab150077) at dilution of 1/1000 (green).
Anti-MAP2 antibody [HM-2] (ab11267) at 1/1000 dilution was used as a counterstain, with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed as the secondary antibody (ab150120) at dilution of 1/1000 (red).
Negative control 1: Anti-Calbindin (ab108404) at 1/250 dilution with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed as the secondary antibody (ab150120) at dilution of 1/1000.
Negative control 2: Anti-MAP2 antibody [HM-2] (ab11267) at 1/200 dilution with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody (ab150077) at dilution of 1/1000.
The nuclear counterstain is DAPI (blue).
Confocal image showing positive staining in subsets of mouse hippocampal primary neuron.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Fluorescence multiplex immunohistochemical analysis of human cerebellum (formalin-fixed paraffin-embedded section). Merged staining of ab300069, GABA A Receptor alpha 6 at 1:500 (1.244 μg/ml) [Panel B], ab239790, DGKZ/DGK-zeta at 1:9000 (0.109 μg/ml) [Panel C], and ab108404, Calbindin at 1:1000 (1.394 μg/ml) [Panel D] on human cerebellum. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab300069, ab239790, and ab108404 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. DAPI (dark blue) was used as a nuclear counter stain.
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All lanes : Anti-Calbindin antibody [EP3478] (ab108404) at 1/2000 dilution (Purified)
Lane 1 : Human fetal brain lysates
Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
Lane 3 : Human kidney lysates
Lane 4 : Mouse brain lysates
Lane 5 : Mouse kidney lysates
Lane 6 : Rat brain lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 30 kDa
Observed band size: 30 kDa -
Immunohistochemistry (Frozen) analysis of mouse cerebellum tissue sections labeling Calbindin with purified ab108404 at 1/500 dilution (3.0 µg/ml). The tissue section was fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. DAPI (blue) was used as nuclear counterstain.
Secondary antibody only control: PBS instead of the primary antibody -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling Calbindin with purified ab108404 at 1/1000 dilution (1.49 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-Calbindin antibody [EP3478] (ab108404) at 1/1000 dilution
Lane 1 : Fetal brain lysate
Lane 2 : Fetal cerebellum lysate
Lane 3 : Fetal hypothalamus lysate
Lane 4 : Fetal pituitary lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 30 kDa -
Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Calbindin with purified ab108404 at 1/150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemistry (Frozen) analysis of rat cerebellum tissue sections labeling Calbindin with purified ab108404 at 1/500 dilution (3.0 µg/ml). The tissue section was fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. DAPI (blue) was used as nuclear counterstain.
Secondary antibody only control: PBS instead of the primary antibody -
Immunohistochemical analysis of mouse frozen cerebellum tissue sections with ab108404 labelling Calbindin at 1/500. Sections were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue). The image shows specific staining on Purkinje cells of mouse cerebellum.
Secondary antibody only was used as negative control and is shown in the inset.
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ab108404 showing positive staining in Normal kidney tissue.
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ab108404, at 1/100 dilution, staining Calbindin in paraffin-embedded Human brain tissue by Immunohistochemistry.
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ab108404 showing negative staining in Normal heart tissue.
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ab108404 showing negative staining in Normal colon tissue.
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ab108404 showing negative staining in Normal liver tissue.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (22)
ab108404 has been referenced in 22 publications.
- Forsthofer M & Straka H Homeostatic plasticity of eye movement performance in Xenopus tadpoles following prolonged visual image motion stimulation. J Neurol 270:57-70 (2023). PubMed: 35947153
- Frontzek K et al. A conformational switch controlling the toxicity of the prion protein. Nat Struct Mol Biol 29:831-840 (2022). PubMed: 35948768
- Yu H et al. Infralimbic medial prefrontal cortex signalling to calbindin 1 positive neurons in posterior basolateral amygdala suppresses anxiety- and depression-like behaviours. Nat Commun 13:5462 (2022). PubMed: 36115848
- Yamasaki S et al. A Genetic modification that reduces ON-bipolar cells in hESC-derived retinas enhances functional integration after transplantation. iScience 25:103657 (2022). PubMed: 35024589
- Yamasaki S et al. Addition of Chk1 inhibitor and BMP4 cooperatively promotes retinal tissue formation in self-organizing human pluripotent stem cell differentiation culture. Regen Ther 19:24-34 (2022). PubMed: 35059477