Description

Size

100 µl

Catalog no.:

AS10 931

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Supplementary 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

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.

Description

Associated membrane protein types are lipopolysaccharide selective barriers. Biological membranes include cell membranes, outer coverings of cells or organelles that allow passage of certain proteins and nuclear membranes, which cover a cell nucleus; and tissue membranes, such as mucosae and serosae. 

Storage condition

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.

Scientific context

AtPCaP1 is a protein associated with plasma membrane with ability to bind Ca, calmodulin and phosphatidylinositol phoshphates. Alternative names: At4g20260, endomembrane-associated protein.

Verified reactivity

Arabidopsis thaliana, Raphanus sativus (41 kDa), Brassica rapa (42 kDa), Brassica rapa var. glabra Regel (43 kDa), Brassica oleracea var. italica (41 kDa)

Connected products

antibodies to plasma membrane proteinsPlant protein extraction buffer Secondary antibodies

Immunogen

KLH-conjugated peptide derived from Arabidopsis thaliana PCaP1 sequence Q96262, At4g20260

No reactivity

no confirmed exceptions from predicted reactivity known in the moment

Notes

Serum contains less than 0.1 % of sodium azide as preservative.

How to reconstitute

For reconstitution add 100 µl of sterile water.

Recommended dilutions for use

1 : 2000 with standard ECL (WB)

Verified applications

western blot (WB)

Molecular weight (expected | аpparent)

24.5 | 36 kDa

Possible reactivity

Brassicaceae

Available ordering format

Lyophilized

Clone

Polyclonal

Clonality

Polyclonal

TAIR number

AT4G20260

Raised in

Rabbit

Protein number

Q96262

Purification

Serum