Calretinin antibody (AA 38-151)

Catalog No. ABIN968244

Product Details anti-Calretinin Antibody

Target: Calretinin (CALB2) (Calbindin 2 (CALB2))

Binding Specificity: AA 38-151

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This Calretinin antibody is un-conjugated

Application: Western Blotting (WB), BioImaging (BI)

Cross-Reactivity: Human, Mouse (Murine)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
3. Triton is a trademark of the Dow Chemical Company.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
6. Please refer to us for technical protocols.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Rat Calretinin aa. 38-151

Clone: 34-Calretinin

Isotype: IgG1

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN967389, ABIN968545

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-Calretinin antibody (ABIN968244)

Chua, Fletcher, Kalloniatis: "Functional remodeling of glutamate receptors by inner retinal neurons occurs from an early stage of retinal degeneration." in: The Journal of comparative neurology, Vol. 514, Issue 5, pp. 473-91, (2009) (PubMed).

Acosta, Bumsted OBrien, Tan, Kalloniatis: "Emergence of cellular markers and functional ionotropic glutamate receptors on tangentially dispersed cells in the developing mouse retina." in: The Journal of comparative neurology, Vol. 506, Issue 3, pp. 506-23, (2008) (PubMed).

Arnold, Heintz: "A calcium responsive element that regulates expression of two calcium binding proteins in Purkinje cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 94, Issue 16, pp. 8842-7, (1997) (PubMed).

Schurmans, Schiffmann, Gurden, Lemaire, Lipp, Schwam, Pochet, Imperato, Böhme, Parmentier: "Impaired long-term potentiation induction in dentate gyrus of calretinin-deficient mice." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 94, Issue 19, pp. 10415-20, (1997) (PubMed).

Schwaller, Durussel, Jermann, Herrmann, Cox: "Comparison of the Ca2+-binding properties of human recombinant calretinin-22k and calretinin." in: The Journal of biological chemistry, Vol. 272, Issue 47, pp. 29663-71, (1997) (PubMed).

Parmentier, Lefort: "Structure of the human brain calcium-binding protein calretinin and its expression in bacteria." in: European journal of biochemistry / FEBS, Vol. 196, Issue 1, pp. 79-85, (1991) (PubMed).

Target Details for Calretinin

Target: Calretinin (CALB2) (Calbindin 2 (CALB2))

Alternative Name: Calretinin (CALB2 Products)

Synonyms: CAB29 antibody, CAL2 antibody, CR antibody, calb2l antibody, wu:fq18e08 antibody, zgc:73115 antibody, calb2 antibody, wu:fq17g09 antibody, zgc:73099 antibody, calbindin 2 antibody, calbindin 2a antibody, calbindin 2b antibody, CALB2 antibody, Calb2 antibody, calb2a antibody, calb2b antibody

Background: A low level of intracellular free Ca2+ is essential for a variety of cellular functions and is mediated by the sequestration of Ca2+ in the ER, by the action of plasma membrane Ca2+ pumps, and by Ca2+ binding proteins. The Ca2+ binding protein, Calretinin, is a member of the calmodulin superfamily whose members promote calcium homeostasis by acting as buffers of intracellular free Ca2+. Although Calretinin is expressed in certain populations of neurons throughout the central and peripheral nervous systems, it is primarily expressed in the olfactory bulb and auditory pathways. Calretinin most closely resembles calbindin D28k, another member of the calmodulin family. These two proteins share 58% amino acid identity and contain six EF-hand Ca2+ binding motifs. Excess Calretinin or calbindin results in decreased transcription from PCE1, a calcium responsive element in the calmodulin II promoter. This supports the role of Calretinin as a regulator of intracellular free Ca2+ levels. Furthermore, truncated forms of Calretinin have been found in certain tumor cells, but are absent from normal cells which express wild type Calretinin. Thus, these isoforms may have a role in tumorigenesis.

Molecular Weight: 29 kDa