Plant/Algal cell biology / Compartment markers / Vacuole

product information

Pathogenesis-related (PR) proteins, are induced in response to the infection of plants with microbial pathogens. Combinations of glucanase I and chitinase I are potent inhibitors of fungal growth in vitro however precise mechanism of that is still not known.  Glucanase I  (PR-2) and chitinase I (PR-3) contribute to defense against fungal infection and are currently used as markers for innate immunity, and in particular the ethylene/jasmonate signalling pathway in pathogenesis.

purified tobacco class I chitinase. The preparation used is a mixture of two class I isoforms (Shinshi et al., 1990; van Buuren et al., 1992): 1) Chitinase A (CHN A) P08252 encoded by gene chn48 derived from the N. tomentosiformis ancestor of tobacco. 2) Chitinase B (CHN B) P24091 encoded by gene chn50 derived from the N. sylvestris ancestor of tobacco.

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.

western blot (WB)

AS07 208 | anti-PR-2 | GLU I | class I beta-1,3-glucanase

antibody is recognizing closely related tobacco class I isoforms: endochitinase A CHN-A (ca. 34 kDa) and endochitinase B CHN-B (ca. 32 kDa)

This antibody can be used as a marker of vacuolar contents Keefe et al. (1990). The effect of ethylene on the cell-type-specific and intracellular localization of β-1,3-glucanase and chitinase in tobacco leave. Plant 182: 43-51. 

application information

8 ug/ml with standard ECL (WB)

35, 34 | 32 and 34 kDa

Nicotiana tabacum

dicots including: Arabidopsis thaliana, Solanum tuberosum, Vitis vinifera, monocots including: Zea mays

no confirmed exceptions from predicted reactivity known in the moment

Important note: For blocking 5 % skim milk in PBS without Ca++ should be used

Sticher et al. (1993). Posttranslational processing of a new class of hydroxyproline-containing proteins: Prolyl hydroxylation and C-terminal cleavage of tobacco (Nicotiana tabacum) vacuolar chitinase. Plant Physiol. 101, 1239-1247.

application example

Detection of tobacco chitinase I in ng loaded per respective well using anti-tobacco chitinase I antibodies. Primary antibodies have been used at 8µg/ml.