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Western blot analysis of extracts from MCF7 cells, untreated or stimulated with IGF-I, using Phospho-IGF-I Receptor β (Tyr1135) (DA7A8) Rabbit mAb (upper) and IGF-I Receptor β Antibody #3027 (lower).
Western blot analysis of extracts from 3T3-L1 adipocytes, untreated or insulin-treated (100 nM for the indicated times), using Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody (upper) or control IR antibody (lower).
Western blot analysis of untreated and IGF-treated Hela cell extracts as well as untreated and insulin-treated H-4-II-E cell extracts using Phospho-IGF-I-Receptor beta (Tyr1135/1136)/Insulin Receptor beta (Tyr1150/1151)(19H7) Rabbit mAb
Western blot analysis of extracts from MCF-7 cells, untreated or IGF-I-treated, using Phospho-IGF-I Receptor β (Tyr980) (C14A11) Rabbit mAb (upper) and IGF-I Receptor β (111A9) Rabbit mAb #3018 (lower).
Immunprecipitation of Insulin Receptor beta from insulin treated mIMCD-3 cell extracts using Insulin Receptor beta antibody (Lane 1) Lane 2: No antibody control. Lane 3: Input control.
Flow cytometric analysis of fixed and permeabilized SK-UT-1 cells (blue, negative) and MCF7 cells (green, positive) using IGF-I Receptor β (D23H3) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from 293 cells, untreated or IGF-I-treated (100 nM for 2 minutes), using Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody (upper) or control IGF-I Receptor antibody (lower).
Phospho-IGF-I Receptor
β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb specifically binds to tyrosine phosphorylated IGF-1 and insulin receptors, but not other phosphorylated tyrosine kinases. Western blot analysis of of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β(Tyr1150/1151) (19H7) Rabbit mAb (upper) or Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (lower).
Western blot analysis of extracts from various cell lines using Insulin Receptor β (4B8) Rabbit mAb.
Confocal immunofluorescent analysis of MCF7 (left) and SK-UT-1 (right) cells using IGF-I Receptor β (D23H3) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of extracts from 293 (IGF-I receptor β+) and SK-UT-1 (IGF-I receptor β-) cells using IGF-I Receptor β (D23H3) XP® Rabbit mAb (upper) or β-Actin Antibody #4967 (lower).
| Product # | Size | Price |
|---|---|---|
| 8338T | 1 Kit (6 x 20 µl) | $ 458 |
| Product Includes | Quantity | Applications | Reactivity | MW(kDa) | Isotype |
|---|---|---|---|---|---|
| Phospho-IGF-I Receptor β (Tyr1135) (DA7A8) Rabbit mAb 3918 | 20 µl |
|
H M R | 95 | Rabbit IgG |
| Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody 3021 | 20 µl |
|
H M R | 95 | Rabbit |
| Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb 3024 | 20 µl |
|
H M R | 95 | Rabbit IgG |
| Phospho-IGF-I Receptor β (Tyr980) (C14A11) Rabbit mAb 4568 | 20 µl |
|
H M R | 95 | Rabbit IgG |
| Insulin Receptor β (4B8) Rabbit mAb 3025 | 20 µl |
|
H M R | 95 | Rabbit IgG |
| IGF-I Receptor β (D23H3) XP® Rabbit mAb 9750 | 20 µl |
|
H M R Mk | 95 | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody 7074 | 100 µl |
|
Goat |
Product Information
The Phospho-Insulin/IGF Receptor Antibody Sampler Kit provides an economical means of evaluating total Insulin Receptor and IGF-I Receptor β protein levels as well as Insulin and IGF-I Receptor β phosphorylated at specific sites. The kit includes enough antibody to perform two western blot experiments with each primary antibody.
| MW (kDa) | 95 |
Each antibody in the Phospho-Insulin/IGF Receptor Antibody Sampler Kit recognizes endogenous levels of the specific target protein, with activation state-specific antibodies recognizing target proteins only when phosphorylated at the indicated residues. IGF-I Receptor ß (D23H3) XP® Rabbit mAb detects endogenous levels of total IGF-I receptor ß protein. This antibody does not cross-react with insulin receptor.Phospho-IGF-I Receptor beta (Tyr1135) (DA7A8) Rabbit mAb detects endogenous levels of IGF-I receptor only when phosphorylated at Tyr1135. This antibody cross-reacts with Tyr1150 of insulin receptor and may also cross-react with other overexpressed related tyrosine-phosphorylated tyrosine kinases. Phospho-IGF-I Receptor ß (Tyr1131)/Insulin Receptor ß (Tyr1146) Antibody detects endogenous levels of Tyr1131-phosphorylated IGF-I receptor and Tyr1146-phosphorylated insulin receptor. This antibody cross-reacts with activated PDGF, FGF and EGF receptors; ErbB2; and c-Met. Phospho-IGF-I Receptor ß (Tyr1135/1136)/Insulin Receptor ß (Tyr1150/1151) (19H7) Rabbit mAb detects endogenous levels of IGF-I receptor and insulin receptor only when phosphorylated at tyrosine 1135/1136 or tyrosine 1150/1151, and does not cross-react with other related tyrosine-phosphorylated tyrosine kinases. Phospho-IGF-I Receptor ß (Tyr980) (C14A11) Rabbit mAb detects endogenous levels of IGF-I receptor ß protein only when phosphorylated at Tyr980, and may cross-react with activated insulin receptors and FLT3. Insulin Receptor ß (4B8) Rabbit mAb detects endogenous levels of total insulin receptor ß and does not cross-react with IGF-I receptor ß.
Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy-terminal residues of human IGF-IR β or residues surrounding Tyr960 of human insulin receptor β. Activation state-specific monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1135 of human IGF-I Receptor β, Tyr1135/1136 of human IGF-I Receptor β, or Tyr980 of human IGF-I Receptor β. Activation state-specific polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues of human IGF-I Receptor β. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Type I insulin-like growth factor receptor (IGF-IR) is a transmembrane receptor tyrosine kinase that is widely expressed in many cell lines and cell types within fetal and postnatal tissues (1-3). Receptor autophosphorylation follows binding of the IGF-I and IGF-II ligands. Three tyrosine residues within the kinase domain (Tyr1131, Tyr1135, and Tyr1136) are the earliest major autophosphorylation sites (4). Phosphorylation of these three tyrosine residues is necessary for kinase activation (5,6). Insulin receptors (IRs) share significant structural and functional similarity with IGF-I receptors, including the presence of an equivalent tyrosine cluster (Tyr1146/1150/1151) within the kinase domain activation loop. Tyrosine autophosphorylation of IRs is one of the earliest cellular responses to insulin stimulation (7). Autophosphorylation begins with phosphorylation at Tyr1146 and either Tyr1150 or Tyr1151, while full kinase activation requires triple tyrosine phosphorylation (8).
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