Monoclonal Anti-Calbindin-D-28K antibody produced in mouse
SIGMA/C9848 - clone CB-955, ascites fluid
Synonym: Calbindin Antibody; Calbindin Antibody - Monoclonal Anti-Calbindin-D-28K antibody produced in mouse
MDL Number: MFCD00164553
Product Type: Chemical
Immunohistochemistry-immunofluorescence JOURNAL CITATION: A mouse model for fucosidosis recapitulates storage pathology and neurological features of the milder form of the human disease. By: Wolf, H., Damme, M., et al. in Dis Model Mech, 2016. PubMed ID: 27491075 Image collected and cropped by CiteAb from the following publication, (http://dmm.biologists.org/lookup/doi/10.1242/dmm.025122), provided under a CC-BY license. Image might reflect a new usage that is not reflected in claims on product description or independently verified by Merck KGaA, Darmstadt, Germany. For Research Use Only. Not for use in diagnostic procedures.
Immunohistochemistry-immunofluorescence JOURNAL CITATION: Renal stromal miRNAs are required for normal nephrogenesis and glomerular mesangial survival. By: Phua, Y. L., Chu, J. Y., et al. in Physiol Rep, 2015. PubMed ID: 26438731 Image collected and cropped by CiteAb from the following publication, (http://doi.wiley.com/10.14814/phy2.12537), provided under a CC-BY license. Image might reflect a new usage that is not reflected in claims on product description or independently verified by Merck KGaA, Darmstadt, Germany. For Research Use Only. Not for use in diagnostic procedures.
Immunohistochemistry-immunofluorescence JOURNAL CITATION: Inhibition of colony-stimulating factor 1 receptor early in disease ameliorates motor deficits in SCA1 mice. By: Qu, W., Johnson, A., et al. in J Neuroinflammation, 2017. PubMed ID: 28545543 Image collected and cropped by CiteAb from the following publication, (http://jneuroinflammation.biomedcentral.com/articles/10.1186/s12974-017-0880-z), provided under a CC-BY license. Image might reflect a new usage that is not reflected in claims on product description or independently verified by Merck KGaA, Darmstadt, Germany. For Research Use Only. Not for use in diagnostic procedures.
Immunofluorescence
Double staining with Rabbit Anti-cfos (Cat. No. F7799) (red) at a 1:5,000 dilution, and Monoclonal Anti-Calbindin-D-28K (Cat. No. C9848) (green) at a 1:400 dilution, reveals neuronal activation in a sub-population of lateral septal cells. Developed with biotinylated Donkey Anti-Rabbit, followed by Streptavidin-Cy3 (Cat. No. S6402) and FITC-labeled Goat Anti-Mouse.
From Dr. S. Shoham, Herzog Hospital Research Lab, Jerusalem, Israel.
Double staining with Rabbit Anti-cfos (Cat. No. F7799) (red) at a 1:5,000 dilution, and Monoclonal Anti-Calbindin-D-28K (Cat. No. C9848) (green) at a 1:400 dilution, reveals neuronal activation in a sub-population of lateral septal cells. Developed with biotinylated Donkey Anti-Rabbit, followed by Streptavidin-Cy3 (Cat. No. S6402) and FITC-labeled Goat Anti-Mouse.
From Dr. S. Shoham, Herzog Hospital Research Lab, Jerusalem, Israel.
Enhanced Validation-By Independent Antibodies Immunohistochemistry. Formalin-fixed, paraffin-embedded Rat Cerebellum sections stained with 1:3,000 Monoclonal Anti-Calbindin-D-28K antibody produced in Mouse, Clone: CB-955 (Cat. No.C9848) (A). The antibody was developed using 20 μg/mL Anti-Mouse IgG (Fab specific)-Biotin antibody produced in goat (Cat. No. B0529) followed by 20 μg/mL ExtrAvidin®-Peroxidase (Cat. No. E2886), and 1:2,000 Anti-Calbindin-D-28K (EG-20) antibody produced in Rabbit (Cat. No. C2724) (B). The antibody was developed using 20 μg/mL Goat Anti-Rabbit IgG (whole molecule)-Biotin (Cat. No. B8895) followed by 20 μg/mL ExtrAvidin®-Peroxidase (Cat. No. E2886). Results Two Anti-Calbindin-D-28K antibodies, C9848 (A) and C2724 (B), target different regions of Calbindin-D-28K show similar staining profiles between the two antibodies, demonstrating Independent Antibody Verification.
Immunohistochemistry Formalin-fixed, paraffin-embedded tissue section of rat cerebellum stained with 1:3,000 Monoclonal Anti-Calbindin-D-28K antibody produced in mouse clone: CB-955 (Cat. No. C9848) followed by Anti-Mouse IgG (Fab specific)-Biotin antibody produced in goat (Cat. No. B7151) and by ExtrAvidin®-Peroxidase (Cat. No. E2886).
Immunoblotting Whole extract of rat leg muscle was separated on SDS-PAGE and probed with Monoclonal Anti-Calbindin-D-28K Clone: CB-955 (Cat. No. C9848). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. Antibody dilution 1:1,500 2. Antibody dilution 1:3,000
| antibody form | ascites fluid |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | CB-955, monoclonal |
| concentration | 15-55 mg/mL |
| conjugate | unconjugated |
| contains | 15 mM sodium azide |
| enhanced validation | independent Learn more about Antibody Enhanced Validation  |
| isotype | IgG1 |
| mol wt | antigen 28 kDa |
| Quality Level | 200 |
| shipped in | dry ice |
| species reactivity | mouse, human, guinea pig, canine, sheep, rat, pig, rabbit, feline, bovine, goat |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunohistochemistry (formalin-fixed, paraffin-embedded sections): 1:3,000 using rat cerebellum sections |
| indirect ELISA: suitable | |
| western blot: 1:3,000 using MDBK cell extract | |
| UniProt accession no. | P05937  |
| Application: | Monoclonal Anti-Calbindin-D-28K antibody produced in mouse may be used in immunohistochemistry (alcohol-, formalin-, and methacarn-fixed, paraffin-embedded, and frozen tissue sections). Enzymatic predigestion of formalin-fixed, paraffin-embedded sections improves immunohistochemical staining. The antibody produced in mouse is suitable for indirect ELISA, western blotting at a working dilution of 1:3000 using MDBK cell extract and indirect immunoperoxidase staining with a working dilution of 1:3000 using trypsin-digested, formalin-fixed, paraffin-embedded rat cerebellum sections. It was used for immunohistochemistry and immunocytochemistry at a working dilution of 1:500 in a study to understand impaired cerebellar development and function in mice lacking CAPS2, a protein involved in neurotrophin release. It was used for immunostaining to gauge the development of neuronal and glial cells in the inner nuclear layer of islet-class factor Islet-1 (Isl1)-null and control retinas. It was used for immunohistochemical colocalization studies to confirm postsynaptic localization of the RGS11 complex in a study. |
| Biochem/physiol Actions: | Calbindin-D-28K is involved in intramembranous transport of calcium or acts as an intracellualr calcium buffer in many CNS structures such as subpopulations of central and peripheral nervous system neurons. It is also present in certain epithelial cells involved in Ca2+ transport such as distal tubular cells and cortical collecting tubules of the kidney, and in enteric neuroendocrine cells. |
| Biochem/physiol Actions: | The antibody does not react with other members of the EF-hand family, such as calbindin-D-9K, calretinin, myosin light chain, parvalbumin, S-100a, S-100b, S-100A2 (S100L) and S-100A6 (calcyclin). A weaker reactivity was observed with chicken calbindin-D-28K. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | Calbindin-D-28K is a highly conserved 28 kDa calcium binding protein with broad tissue distribution. Monoclonal Anti-Calbindin-D-28K (mouse IgG1 isotype) is derived from the CB-955 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB/c mice immunized with a purified bovine kidney calbindin-D-28K. The isotype is determined by a double diffusion immunoassay using Mouse Monoclonal Antibody Isotyping Reagents, Catalog Number ISO2. Calbindin-D-28K belongs, together with calmodulin, S-100, parvalbumin, troponin C and other proteins, to a family of low molecular weight calcium-binding proteins (CaBPs). These CaBPs have homologous primary structures, which contain polypeptide folds of the EF-hand type for the acceptance of Ca2+. Calbindin is an attractive neuroanatomical marker. Despite its broad tissue distribution, calbindin-D-28K exhibits a cell-type specific expression pattern. Thus, it has been immunocytochemically localized in selected cells in many CNS structures, where it is thought to act either as an intracellular calcium buffer, or in the intramembranous transport of calcium. It is found predominantly in subpopulations of central and peripheral nervous system neurons, and in certain epithelial cells involved in Ca2+ transport such as distal tubular cells and cortical collecting tubules of the kidney, and in enteric neuroendocrine cells. Monoclonal antibody reacting specifically with calbindin-D-28K is instrumental in the localization of calbindin in different tissues and cell populations. |
| Immunogen: | Bovine kidney calbindin-D |
| Physical form: | Reagent is supplied as ascites fluid with 15mM sodium azide as a preservative. |
| Preparation Note: | For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing, or storage in "frost free" freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |