Agrisera/AtPCaP1 | Arabidopsis thaliana plasma membrane cationbinding protein1/AS10 931/


product information
Background		AtPCaP1 is a protein associated with plasma membrane with ability to bind Ca, calmodulin and phosphatidylinositol phoshphates. Alternative names: At4g20260, endomembrane-associated protein.
Immunogen		KLH-conjugated peptide derived from Arabidopsis thaliana PCaP1 sequence Q96262, At4g20260
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Serum
Format		Lyophilized
Quantity		100 µl
Reconstitution		For reconstitution add 100 µl of sterile water.
Storage		store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications		western blot (WB)
Related products		antibodies to plasma membrane proteins Plant protein extraction buffer Secondary antibodies
Additional information		Serum contains less than 0.1 % of sodium azide as preservative.

application information
Recommended dilution		1 : 2000 with standard ECL (WB)
Expected \| apparent MW		24.5 \| 36 kDa
Confirmed reactivity		Arabidopsis thaliana, Raphanus sativus (41 kDa), Brassica rapa (42 kDa), Brassica rapa var. glabra Regel (43 kDa), Brassica oleracea var. italica (41 kDa)
Predicted reactivity		Brassicaceae
Not reactive in		no confirmed exceptions from predicted reactivity known in the moment
Additional information		It is confirmed that the nock-out mutant (SALK_022955, Col-0 background) has no mRNA or protein of PCaP1. In the immunoblot, crude membrane fractions prepared from wild-type and pcap1 knock-out mutant plants were examined. Therefore the antibody to PCaP1 specifically recognizes PCaP1 and can be used for immunoblotting. For application in immunocytochemistry cross reacting band in proximity of Rubisco has to be removed by incubation of an antibody with a part of memrane with background band. Protocol for preparation of crude membranes This protocol will help to prepare crude membrane fractions from very small amount of plant tissue. (1) Homogenize roots or root tips with the buffer without SDS. (2) Centrifuge the homogenate in Eppendorf tubes at 2 000 rpm for 2 min to remove cell wall, nuclei, plastids and mitochondria. (3) Centrifuge the obtained supernatant at 30 000 g for 20 min to separate soluble components. Please use adequate tubes for ultracentrifugation. (4) Suspend the obtained precipitate (crude membranes) with SDS buffer with DTT or beta-mercaptoethanol. (5) Heat the sample solution obtained the step (4) at 70 C for 5 min. (6) Apply the denatured sample to SDS-PAGE. Courtesy Prof. Msayoshi Maeshima, Nagoya University, Japan
Selected references		Ide et al. (2007) Molecular properties of a novel, hydrophilic cation-binding protein associated with the plasma membrane. J. Exp. Botany, 58: 1173-1183. Nagasaki et al. (2008) A hydrophilic cation-binding protein of Arabidopsis thaliana, AtPCaP1, is localized to plasma membrane via N-myristoylation and interacts with calmodulin and the posphatidylinositol phosphates PtdIns(3,4,5)P3 and PtdIns(3,5)P2. FEBS J. 275: 2267-2282.

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Agrisera/AtPCaP1 | Arabidopsis thaliana plasma membrane cationbinding protein1/AS10 931/