Product Pathways - PathScan ELISA
PathScan® Total Insulin Receptor β Sandwich ELISA Kit #7069
|7069S||1 Kit (96 assays)||---||In Stock||---|
|7069||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
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|Kit Includes||Volume||Solution Color|
|Insulin Receptor beta Mouse Antibody coated microwells||96 tests|
|Insulin Receptor beta Rabbit Detection Antibody||11 ml||Green|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
PathScan® Total Insulin Receptor β Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected levels of Insulin Receptor β protein. An Insulin Receptor β Mouse mAb has been coated on the microwells. After incubation with cell lysates, Insulin Receptor β protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, an Insulin Receptor β Rabbit mAb is added to detect captured Insulin Receptor β protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Insulin Receptor β protein.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Total Insulin Receptor β Sandwich ELISA Kit #7069 detects transfected levels of human Insulin Receptor β protein, as shown in Figure 1. 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.
ELISA - Western correlation
Figure 1: Treatment of CHO-IR/IRS-1 cells with insulin stimulates tyrosine phosphorylation of Insulin Receptor β which is detected by PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit #7082 (top, right). Similar levels of Insulin Receptor β protein from both nonphospho or phospho lysates are detected by PathScan® Total Insulin Receptor β Sandwich ELISA Kit #7069 (top, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb #3024 (right panel) or Insulin Receptor β (4B8) Rabbit mAb #3025 (left panel) are shown in the bottom figure. CHO-IR/IRS-1 cells stably overexpress the tranfected human insulin receptor and rat IRS-1.
Insulin receptor (INSR) is a membrane receptor tyrosine kinase. The receptor molecule consists of a disulfide linked heterodimer. The α subunit is a 135 kDa extracellular fragment, and the β subunit is a 95 kDa fragment containing an extracellular domain, a single transmembrane domain, and an intracellular tyrosine kinase domain (1). Insulin ligand binding to this receptor results in receptor autophosphorylation and tyrosine kinase activation. INSR catalyzes the tyrosine phosphorylation of molecules such as IRS, Gab1, Shc, and Cbl, which further activate the downstream MAPK, PI3K, and TC10 pathways. This eventually leads to increases in glucose uptake and metabolism as well as cell growth (2,3). INSR has peptide substrate specificity similar to other receptor tyrosine kinase members, preferring acidic residues at the -1 to -4 positions and large hydrophobic amino acids at positions +1 and +3 (4).
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- 7254 PathScan® Phospho-Insulin Receptor β (Tyr1146) Sandwich ELISA Kit
- 7258 PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit
- 7326 PathScan® Phospho-Insulin Receptor β (Tyr1345) Sandwich ELISA Kit
- 7082 PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit
- 3025 Insulin Receptor β (4B8) Rabbit mAb
- 3020 Insulin Receptor β (L55B10) Mouse mAb
- 3026 Phospho-Insulin Receptor β (Tyr1345) (14A4) Rabbit mAb
- 3023 Phospho-Insulin Receptor β (Tyr1361) (84B2) Rabbit mAb
- 3024 Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.