Product Pathways - Tyrosine Kinase / Adaptors
PathScan® Phospho-HER4/ErbB4 (panTyr) Sandwich ELISA Kit #13125
|13125S||1 Kit (96 assays)||---||In Stock||---|
|13125||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|
|HER4/ErbB4 Rabbit mAb coated microwells||96 tests|
|Phospho-Tyrosine Mouse Detection Antibody||11 ml||Green|
|Anti-mouse 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).
The PathScan® Phospho-HER4/ErbB4 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-HER4/ErbB4 (panTyr) protein. A HER4/ErbB4 rabbit mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-HER4/ErbB4 proteins are captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse antibody is added to detect captured tyrosine-phosphorylated HER4/ErbB4 proteins. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of HER4/ErbB4 phosphorylated on tyrosine residues.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Phospho-HER4/ErbB4 (panTyr) Sandwich ELISA Kit #13125 detects endogenous levels of tyrosine-phosphorylated HER4/ErbB4 protein in human cells, 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 T-47D cells with Human Neuregulin-1 (hNRG-1) #5218 stimulates tyrosine phosphorylation of HER4/ErbB4 protein, as detected by PathScan® Phospho-HER4/ErbB4 (panTyr) Sandwich ELISA Kit #13125, but does not affect the level of total HER4/ErbB4 detected by PathScan® Total HER4/ErbB4 Sandwich ELISA Kit #13010. The absorbance readings at 450 nm are shown in the top figure while corresponding western blots using HER4/ErbB4 (111B2) Rabbit mAb #4795 (left panel) and Phospho-HER4/ErbB4 (Tyr984) Antibody #3790 (right panel) are shown in the bottom figure.
Figure 2: The relationship between protein concentration of lysates from untreated and hNRG-1-treated T-47D cells and the absorbance at 450 nm as detected by the PathScan® Phospho-HER4/ErbB4 (panTyr) Sandwich ELISA Kit is shown. Starved T-47D cells (85% confluence) were treated with Human Neuregulin-1 (hNRG-1) #5218 (10 ng/ml) for 2-5 min at 37°C and then lysed.
Research studies have implicated the HER/ErbB receptor tyrosine kinase family in normal development, cardiac function and cancer (1,2). HER4/ErbB4, like other family members, has four ectodomains, a single transmembrane domain and a cytoplasmic tail containing the active tyrosine kinase domain (3). By binding to neuregulins and/or EGF family ligands, ErbB4 forms either a homodimer or heterodimer with other ErbB family members, which results in receptor activation and signaling (3). ErbB4 is ubiquitously expressed with the highest expression occurring in brain and heart. The expression of ErbB4 in breast cancer, pediatric brain cancer and other types of carcinomas has been reported in research studies suggesting that ErbB4 expression is involved in both normal tissue development and carcinogenesis (3).
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- 13010 PathScan® Total HER4/ErbB4 Sandwich ELISA Kit
- 4795 HER4/ErbB4 (111B2) Rabbit mAb
- 4757 Phospho-HER4/ErbB4 (Tyr1284) (21A9) Rabbit mAb
- 3790 Phospho-HER4/ErbB4 (Tyr984) Antibody
- 7076 Anti-mouse 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.