Revision 4

#57606

Store at +4C

PathScan® RP Total ULK1 Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H M

UniProt ID:
#O75385

Entrez-Gene Id:
#8408

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color Storage Temp
ULK1 Rabbit mAb Coated Microwells9707896 tests+4C
ULK1 Rabbit Detection mAb257421 eaRed (Lyophilized)+4C
HRP Diluent135155.5 mlRed+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Wash Buffer (20X)980125 ml+4C
Cell Lysis Buffer (10X)980315 ml-20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The rapid protocol (RP) PathScan® RP Total ULK1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of ULK1 protein in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with ULK1 in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of ULK1. Learn more about all of your ELISA kit options here.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

The PathScan® RP Total ULK1 Sandwich ELISA Kit detects endogenous levels of ULK1 protein. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Two related serine/threonine kinases, UNC-51-like kinase 1 and 2 (ULK1, ULK2), were discovered as mammalian homologs of the C. elegans gene unc-51 in which mutants exhibited abnormal axonal extension and growth (1-4). Both proteins are widely expressed and contain an amino-terminal kinase domain followed by a central proline/serine rich domain and a highly conserved carboxy-terminal domain. The roles of ULK1 and ULK2 in axon growth have been linked to studies showing that the kinases are localized to neuronal growth cones and are involved in endocytosis of critical growth factors, such as NGF (5). Yeast two-hybrid studies found ULK1/2 associated with modulators of the endocytic pathway, SynGAP, and syntenin (6). Structural similarity of ULK1/2 has also been recognized with the yeast autophagy protein Atg1/Apg1 (7). Knockdown experiments using siRNA demonstrated that ULK1 is essential for autophagy (8), a catabolic process for the degradation of bulk cytoplasmic contents (9,10). It appears that Atg1/ULK1 can act as a convergence point for multiple signals that control autophagy (11), and can bind to several autophagy-related (Atg) proteins, regulating phosphorylation states and protein trafficking (12-16).

Applications Key

ELISA: ELISA

Cross-Reactivity Key

H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

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Figure 1. The absorbance at 450 nm using the PathScan® RP Total ULK1 Sandwich ELISA Kit #57606 for 293T cell extracts (0.1 mg/mL) that were transfected with an expression construct encoding mouse ULK1 (mULK1) or mock transfected is shown in the upper figure. The corresponding western blots using ULK1 antibody (left panel) and β-Actin antibody (right panel) are shown in the lower figure.
ELISA Image 1: PathScan<sup>®</sup> RP Total ULK1 Sandwich ELISA Kit
Figure 2. Treatment of A-172 cells with the mTOR inhibitor Torin 1 inhibits phosphorylation of ULK1 at Ser757 but does not affect the level of total ULK1. The relationship between lysate protein concentration from untreated and Torin 1-treated A-172 cells and the absorbance at 450 nm using the PathScan® RP Total ULK1 Sandwich ELISA Kit #57606 is shown in the upper figure. The corresponding western blots using phospho-ULK1 (Ser757) antibody (left panel) and ULK1 antibody (right panel) are shown in the lower figure. A-172 cells were treated with Torin 1 (250 nM) for 5 hours at 37°C and then lysed.
ELISA Image 2: PathScan<sup>®</sup> RP Total ULK1 Sandwich ELISA Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.