Western blot analysis was performed using whole cell extracts (30 µg) of Mouse Brain (Lane 1), Rat Brain (Lane 2), IMR-32 (Lane 3), SH-SY5Y (Lane 4) and U-2 OS (Lane 5).The blots were probed with Anti-Calbindin D-28K Rabbit Polyclonal Antibody (Product # PA1-931, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4µg/mL 1:2500 dilution). A ~28 kDa band corresponding to Calbindin D-28K was observed across tissues and cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||28 kDa calbindin-D protein purified from rat kidney.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 15mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1 ug/ml|
|Immunoprecipitation (IP)||5 ug/ml|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Blocking Assay (BLOCK)||See 1 publications below|
PA1-931 detects calbindin D-28k protein in rat samples.
PA1-931 has successfully been used in Western blot , ELISA, immunoprecipitation and immunohistochemical procedures. By Western blot, this antibody detects a 28 kDa protein representing calbindin D from rat cerebellum and rat kidney extract. Immunohistochemical staining on paraffin sections demonstrates that calbindin D is localized to the cytoplasm in epithelial cells of distal tubules in human kidney cortex after staining with PA1-931.
The PA1-931 immunizing protein corresponds to the purified 28 kDa calbindin-D protein from rat kidney.
Vitamin D-dependent 28 kDa calcium-binding protein (calbindin D 28k or CB) is a member of the calmodulin superfamily of calcium binding proteins discovered in 1966 by Wasserman and Taylor. The protein has a distinct distribution in the brain and sensory system and is abundant in specific neuronal cell types. Calbindin D-28k constitutes as much as 0.1-1.5 % of the total soluble protein in brain and it may be present at intracellular concentrations up to 1 mM.
Several recent studies have shown that calbindin D can protect cells from degeneration and ischemic injury under extracellular stress. In particular, this is demonstrated in the brain of Alzheimer and quote;s disease patients. Cells containing calbindin D have a considerably lower plaque and tangles burden than those lacking this 28 kDa protein.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Anti-Yo antibody uptake and interaction with its intracellular target antigen causes Purkinje cell death in rat cerebellar slice cultures: a possible mechanism for paraneoplastic cerebellar degeneration in humans with gynecological or breast cancers.
PA1-931 was used in blocking or activating experiment to investigate anit-Yo antibodies in Purkinje cell death
|Greenlee JE,Clawson SA,Hill KE,Wood B,Clardy SL,Tsunoda I,Carlson NG||PloS one (10:null)||2015|