WB
Western blot analysis of Phospho-AS160 (T642) on different lysates with Rabbit anti-Phospho-AS160 (T642) antibody at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 25 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of Phospho-AS160 (T642) on different lysates with Rabbit anti-Phospho-AS160 (T642) antibody at 1/1,000 dilution. Lane 1: PC-12 cell lysate Lane 2: PC-12 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 25 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of Phospho-AS160 (T642) on different lysates with Rabbit anti-Phospho-AS160 (T642) antibody at 1/1,000 dilution. Lane 1: HEK-293 cell lysate Lane 2: HEK-293 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate Lane 3: HEK-293 cell lysate, the membrane treated with λpp for 1 hour Lane 4: HEK-293 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/100,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells starved overnight then treated with or without 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes labeling Phospho-AS160 (T642) with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.ICC/IF
Immunocytochemistry analysis of 293T cells starved overnight then treated with or without 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes labeling Phospho-AS160 (T642) with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Product Name | Phospho-AS160 (T642) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phosphopeptide corresponding to residues surrounding Thr642 of human AS160 protein. |
| Modification | p-T642 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFWB |
| WB:1:1000 ICC/IF:1:100 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | TBC1D4 |
|---|---|
| Gene Synonyms | AS160 NIDDM5 |
| Gene Full Name | TBC1 domain family member 4 |
| Gene Summary | This gene is a member of the Tre-2/BUB2/CDC16 domain family. The protein encoded by this gene is a Rab-GTPase-activating protein, and contains two phopshotyrosine-binding domains (PTB1 and PTB2), a calmodulin-binding domain (CBD), a Rab-GTPase domain, and multiple AKT phosphomotifs. This protein is thought to play an important role in glucose homeostasis by regulating the insulin-dependent trafficking of the glucose transporter 4 (GLUT4), important for removing glucose from the bloodstream into skeletal muscle and fat tissues. Reduced expression of this gene results in an increase in GLUT4 levels at the plasma membrane, suggesting that this protein is important in intracellular retention of GLUT4 under basal conditions. When exposed to insulin, this protein is phosphorylated, dissociates from GLUT4 vesicles, resulting in increased GLUT4 at the cell surface, and enhanced glucose transport. Phosphorylation of this protein by AKT is required for proper translocation of GLUT4 to the cell surface. Individuals homozygous for a mutation in this gene are at higher risk for type 2 diabetes and have higher levels of circulating glucose and insulin levels after glucose ingestion. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Aug 2015] |
| Alternative Names | Acrg embryonic lethality (mouse) minimal region ortholog antibody Acrg embryonic lethality minimal region ortholog antibody Acrg embryonic lethality mouse minimal region ortholog antibody Akt substrate of 160 kDa antibody AS 160 antibody AS160 antibody BUB2 antibody CDC16 antibody KIAA0603 antibody NIDDM5 antibody Acrg embryonic lethality (mouse) minimal region ortholog antibody Acrg embryonic lethality minimal region ortholog antibody Acrg embryonic lethality mouse minimal region ortholog antibody Akt substrate of 160 kDa antibody AS 160 antibody AS160 antibody BUB2 antibody CDC16 antibody KIAA0603 antibody NIDDM5 antibody TBC (Tre 2 BUB2 CDC16) domain containing protein antibody TBC Tre 2 BUB2 CDC16 domain containing protein antibody TBC1 D4 antibody TBC1 domain family member 4 antibody Tbc1d4 antibody TBCD4_HUMAN antibody Tre-2 antibody |
| Molecular Weight(MW) | 147 kDa(Observed band size: 160 kDa) |
| Cellular Localization | Cytoplasm. |
WB
Western blot analysis of Phospho-AS160 (T642) on different lysates with Rabbit anti-Phospho-AS160 (T642) antibody at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 25 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of Phospho-AS160 (T642) on different lysates with Rabbit anti-Phospho-AS160 (T642) antibody at 1/1,000 dilution. Lane 1: PC-12 cell lysate Lane 2: PC-12 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 25 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of Phospho-AS160 (T642) on different lysates with Rabbit anti-Phospho-AS160 (T642) antibody at 1/1,000 dilution. Lane 1: HEK-293 cell lysate Lane 2: HEK-293 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate Lane 3: HEK-293 cell lysate, the membrane treated with λpp for 1 hour Lane 4: HEK-293 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/100,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells starved overnight then treated with or without 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes labeling Phospho-AS160 (T642) with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
ICC/IF
Immunocytochemistry analysis of 293T cells starved overnight then treated with or without 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes labeling Phospho-AS160 (T642) with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-AS160 (T642) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Application Notes | WB:1:1000 ICC/IF:1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
