Calbindin D-28 Antibody

CAB27, CALB1, Calbindin D28, D-28K, Vitamin D-dependent calcium-binding protein
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Price: $350.00
Product ID : 24427

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Description

The antibody has a proven maximum biotin-streptavidin/HRP staining at a 1/5,000 - 1/10,000 dilution in rat striatum, cortex, and hippocampus.

The antiserum has been characterized as specific to calbindin D-28k; please see reference listed below. Optimal dilution will vary depending upon fixation, labeling technique and/or detection system; therefore, a dilution series is recommended.

Photo Description: IHC image of neurons staining for the calbindin in the rat amygdala (above) and the rat cortex (below). The tissue was fixed with 4% formaldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:10000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen. The section was then mounted on slides with permount.   

Calbindin cortex #24427

Host: Rabbit

Quantity / Volume: 100 µL

State: Lyophilized Whole Serum

Reacts With: Mouse, Rat

Availability: In Stock

Alternate Names:  Calbindin; nCAB27; Calbindin D28; D-28K; Vitamin D-dependent calcium-binding protein; Vitamin D-dependent calcium-binding protein, avian-type; CALB1; calbindin 1, 28kDa, anti-Calbindin

RRID:  AB_572222

Immunogen: Bovine Calbindin D-28K

Gene Symbol: CALB1

Entrez Gene ID: 539930

NCBI Gene Aliases: MGC13895

Reviews

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Reviewed by djbarker
10/02/2012 - 10:35:13 AM
Core/Shell Differentiation

Our lab was able to use this for a beautiful differentiation of the Accumbens core and shell.

The protocol followed was essentially identical to the other review written for this product. However, we found that a dilution of 1:5000 was best for our purposes. It should be noted that we use a potassium ferrocyanide counterstain that seems to lighten Calbindin staining. Thus, my pilot studies would suggest that a dilution of 1:7000 would be sufficient for those that are not using such a counterstain.

The differences between 1:5000 and 1:7000 dilutions (as tested in increments of 500) were quite subtle, but did make some difference when trying to find the medial edge of the shell under the microscope. I would definitely recommend trying a range of dilutions for those attempting the same accumbens staining. Also, the DAB reaction was quite fast for the calbindin. We only needed 2-4 minutes before it seemed the reaction had reaches asymptote. Feel free to email me for a full protocol.

David Barker
http://rutgers.academia.edu/DavidBarker
Reviewed by droot
07/10/2012 - 10:07:28 PM
Visualization of dorsolateral ventral pallidum

For our manuscript, Differential roles of ventral pallidum subregions during cocaine self-administration behaviors, in press at Journal of Comparative Neurology, we used the Immunostar rabbit anti-calbindin-d28k primary antibody to visualize the dorsolateral ventral pallidum subregion.

Rats were perfused with saline followed by 4% PF, stored in 30% sucrose, and coronally sectioned to 40 um. We followed the protocols of Zahm/Heimer and colleagues when they discovered the ventral pallidum subregions and their afferent/efferent projection patterns.

The protocol consisted of the following steps:
1. Wash in 0.1M phosphate buffer (PB; pH 7.4)
2. 15 min in 1% sodium borohydride
3. Wash
4. 1 hour blocking with PB containing 0.1% Triton X-100 and 3% normal goat serum
5. Primary antibody overnight at 4C - ImmunoStar rabbit anti-calbindin-d28k diluted 1 : 6000 in PB containing 0.1% Triton X-100 and 3% normal goal serum
6. Wash
7. 1 hour anti-rabbit biotinylated secondary (Vector) 1 : 200 in PB with 0.1% Triton X-100
8. Wash
9. 1 hour ABC (Vector) 1 : 200 in PB with 0.1% Triton X-100
10. Wash
11. Develop with 0.05% DAB for 6 min

Our study involved recording neurons within the ventral pallidum subregions during specific aspects of intravenous cocaine self-administration (approaching toward, responding on, or retreating away from a cocaine-reinforced operandum). In order to verify the placement of microwires within the VP subregions, prior to perfusion we passed current through each stainless steel microwire to leave an iron deposit at the uninsulated tip. After DAB (brown reaction) and mounting, we visualized the iron deposit by incubating in a 5% potassium ferrocyanide and 10% HCl solution (leaving a blue-green reaction).

For our purposes, this antibody strongly labeled fibers in dorsolateral VP with low background. Individual neurons and other fibers were clearly observed within distinct cortical layers that were not involved in our study.

David H. Root, Ph.D.
Rutgers University

Publications

PROD # ANTIBODY CITATION SPECIES REACTIVITY APPL METHOD RESEARCH AREA # CITED BY PUBMED ID
24427 Calbindin D-28K Antibody Kralic, Jason E., et al. "Compensatory alteration of inhibitory synaptic circuits in cerebellum and thalamus of ??aminobutyric acid type A receptor ?1 subunit knockout mice." Journal of Comparative Neurology 495.4 (2006): 408-421. Mouse IHC, IF 60 16485284
24427 Calbindin D-28K Antibody Kuang, Yi, et al. "Dicer1 and MiR-9 are required for proper Notch1 signaling and the Bergmann glial phenotype in the developing mouse cerebellum." Glia 60.11 (2012): 1734-1746. Mouse IF 2 22836445
24427 Calbindin D-28K Antibody Arabadzisz, Dimitrula, et al. "Epileptogenesis and chronic seizures in a mouse model of temporal lobe epilepsy are associated with distinct EEG patterns and selective neurochemical alterations in the contralateral hippocampus." Experimental neurology 194.1 (2005): 76-90. Mouse 57 15899245
24427 Calbindin D-28K Antibody J Comp Neurol. 2007 Jun 20;502(6):990-1002. GABAergic phenotype of periglomerular cells in the rodent olfactory bulb. Panzanelli P1, Fritschy JM, Yanagawa Y, Obata K, Sassoè-Pognetto M. Mouse IF 17444497
24427 Calbindin D-28K Antibody Barker, David J., et al. "Sensitivity to self-administered cocaine within the lateral preoptic IHC 0 24604249
24427 Calbindin D-28K Antibody Murray, Ryan C., et al. "Striatal patch compartment lesions alter methamphetamine-induced behavior and immediate early gene expression in the striatum, substantia nigra and frontal cortex." Brain Structure and Function (2013): 1-17. 0 23625147
24427 Calbindin D-28K Antibody Coffey, Kevin R., et al. "Electrophysiological evidence of alterations to the nucleus accumbens and dorsolateral striatum during chronic cocaine self‐administration." European Journal of Neuroscience (2015). 0 not defined
24427 Calbindin D-28K Antibody Barker, David J., et al. "Sensitivity to self-administered cocaine within the lateral preoptic–rostral lateral hypothalamic continuum." Brain Structure and Function 220.3 (2015): 1841-1854. 1 24604249
24427 Calbindin D-28K Antibody Murray, Ryan C., et al. "Striatal patch compartment lesions alter methamphetamine-induced behavior and immediate early gene expression in the striatum, substantia nigra and frontal cortex." Brain Structure and Function 219.4 (2014): 1213-1229. 4 23625147

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MSDS

This product contains the preservative sodium azide. The concentration percent of the sodium azide is ≤ .09%. Although this hazardous substance is a concentration below that required for the preparation of a Material Safety Data Sheet, we created a standard MSDS for your records.

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