Human Wee1 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Wee1 in Cell Lysate samples.
Colorimetric
Cell Lysate
Sandwich (quantitative)
Human
0.102 - 6.5 ng/mL
1h 30m
= 19 ng/mL
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Acts as a negative regulator of entry into mitosis (G2 to M transition) by protecting the nucleus from cytoplasmically activated cyclin B1-complexed CDK1 before the onset of mitosis by mediating phosphorylation of CDK1 on 'Tyr-15'. Specifically phosphorylates and inactivates cyclin B1-complexed CDK1 reaching a maximum during G2 phase and a minimum as cells enter M phase. Phosphorylation of cyclin B1-CDK1 occurs exclusively on 'Tyr-15' and phosphorylation of monomeric CDK1 does not occur. Its activity increases during S and G2 phases and decreases at M phase when it is hyperphosphorylated. A correlated decrease in protein level occurs at M/G1 phase, probably due to its degradation.
Wee1-like protein kinase, WEE1hu, Wee1A kinase, WEE1
Human Wee1 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Wee1 in Cell Lysate samples.
Wee1-like protein kinase, WEE1hu, Wee1A kinase, WEE1
Colorimetric
Cell Lysate
Sandwich (quantitative)
Human
0.102 - 6.5 ng/mL
1h 30m
Pre-coated microplate (12 x 8 well strips)
= 19 ng/mL
97%
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Extract | n 8 | mean - | SD - | C.V. 2.4 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Extract | n 3 | mean - | SD - | C.V. 2.3 |
Sample type | Average % | Range |
---|---|---|
Sample type Cell Lysate | Average % = 97 | Range 94 - 99 % |
Blue Ice
+4°C
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Human Wee1 SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Wee1 protein in Cell Lysate. Quantitate Human Wee1 with 19 ng/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Wee1 also known as Wee1-like protein kinase or WEE1 serves as an important regulator of cell cycle progression. Wee1 is a protein kinase with a molecular mass of approximately 96 kDa involved in the regulation of the cell cycle by inhibiting the entry into mitosis through phosphorylation of cyclin-dependent kinase 1 (CDK1). Expression of Wee1 occurs throughout various tissues but it is especially important in those that require tight control over cell division like the brain and reproductive organs. By suppressing premature mitosis Wee1 ensures cells have adequate time for DNA repair and completion of critical processes before cell division.
The function of Wee1 extends to its role in maintaining genomic stability. Wee1 operates as part of a regulatory complex and its inhibition results in defective cell cycle arrest potentially leading to DNA damage. The kinase acts to prevent transitions from the G2 to M phase of the cell cycle ensuring cells repair damaged DNA before division. In the context of DNA replication stress Wee1 cooperates with other regulators such as Chk1 to mediate cell cycle arrest therefore safeguarding genomic integrity.
The role of Wee1 manifests significantly within the DNA damage checkpoint pathway and the cell cycle control pathway. In the DNA damage checkpoint pathway Wee1 collaborates with other cell cycle regulators such as ATR and Chk1 to control the cell cycle in response to DNA damages. Wee1's influence on the cell cycle pathway also intersects with CDK1 and Cyclin B where Wee1 modulates the activity of these proteins to control cell cycle transitions. This regulatory action allows cells to coordinate DNA repair and replication with cell division events.
Wee1's regulatory functions relate closely to cancer and neurological disorders. Overexpression or mutation of Wee1 is associated with various cancers including gliomas and breast cancer where it influences cell proliferation by controlling the activity of CDK1. Wee1's relationship with cancer extends to its interactions with p53 and Chk1 proteins both of which are critical in cancer biology. Additionally anomalies in Wee1 expression or function also associate with certain neurological disorders where it may alter cell cycle dynamics and influence neural cell fate under stress conditions.
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Example of human Wee1 standard curve. Background-subtracted data values (mean +/- SD) are graphed.
Interpolated concentration of native Wee1 was measured in duplicate at different sample concentrations. Undiluted samples are 500 µg/mL Raji cell extract. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR.
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