Source 1primary paper2002The Plant Cell
Toolkit/in-gel assay
in-gel assay
Also known as: in-gel assays
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The in-gel assay is a functional assay method used to detect protein kinase activity within a gel matrix. In the cited Arabidopsis study, it was used to show that OST1 is an abscisic acid (ABA)-activated protein kinase involved in stomatal signaling.
Usefulness & Problems
Why this is useful
This assay is useful for functionally detecting kinase activity rather than inferring it only from sequence or expression. In the cited work, it provided evidence that OST1 is activated by ABA, supporting its role in guard-cell ABA signaling.
Problem solved
The assay helps resolve whether a candidate signaling protein exhibits stimulus-dependent kinase activity. In this case, it addressed whether OST1 functions as an ABA-activated protein kinase in Arabidopsis.
Problem links
Need conditional control of signaling activity
DerivedThe in-gel assay is a functional assay method used to detect protein kinase activity within a gel matrix. In the cited Arabidopsis study, it indicated that OST1 is an abscisic acid (ABA)-activated protein kinase involved in stomatal signaling.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Techniques
Functional AssayTarget processes
signalingImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
The available evidence indicates only that kinase activity was assessed by an in-gel assay and that ABA-dependent activation of OST1 was detected. The supplied material does not describe gel composition, embedded substrate, detection chemistry, sample preparation, or required cofactors.
The supplied evidence only supports use of the assay for detecting OST1 kinase activity in one Arabidopsis study. No additional details are provided here on substrate composition, sensitivity, quantitation, or validation across other kinases or organisms.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
Applied hydrogen peroxide or calcium elicits the same degree of stomatal closure in ost1 mutants as in wild type, suggesting OST1 acts between ABA perception and ROS production.
applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is expressed in stomatal guard cells and vascular tissue.
The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue.
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
ost1 mutations do not affect stomatal regulation by light or CO2, supporting a specific role for OST1 in ABA signaling.
the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling
Approval Evidence
1 source1 linked approval claimfirst-pass slug in-gel-assay
In-gel assays indicated that OST1 is an ABA-activated protein kinase
Source:
molecular functionsupports
OST1 is an ABA-activated protein kinase related to Vicia faba AAPK.
In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK).
Source:
Comparisons
Source-backed strengths
The cited evidence shows that the assay directly indicated ABA-activated kinase activity for OST1. This provides functional support for assigning OST1 a molecular role in ABA-responsive stomatal signaling.
Compared with IRAP-pHluorin translocation assay
in-gel assay and IRAP-pHluorin translocation assay address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling
Strengths here: looks easier to implement in practice.
Compared with multicomponent, ligand-functionalized microarrays
in-gel assay and multicomponent, ligand-functionalized microarrays address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling
in-gel assay and root-specific transcriptomic dataset comparison for ethylene responses address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling
Ranked Citations
- 1.