Phospho-Tuberin/TSC2 (Thr1462) Antibody - #AF3334

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Product: Phospho-Tuberin/TSC2 (Thr1462) Antibody
Catalog: AF3334
Description: Rabbit polyclonal antibody to Phospho-Tuberin/TSC2 (Thr1462)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep
Mol.Wt.: 200 kDa; 201kD(Calculated).
Uniprot: P49815
RRID: AB_2834749

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MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(83%), Bovine(100%), Horse(100%), Sheep(100%)
Clonality:
Polyclonal
Specificity:
Phospho-Tuberin/TSC2 (Thr1462) Antibody detects endogenous levels of Tuberin/TSC2 only when phosphorylated at Threonine 1462.
RRID:
AB_2834749
Cite Format: Affinity Biosciences Cat# AF3334, RRID:AB_2834749.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

FLJ43106; LAM; OTTHUMP00000158940; OTTHUMP00000198394; OTTHUMP00000198395; PPP1R160; Protein phosphatase 1, regulatory subunit 160; tsc2; TSC2_HUMAN; TSC4; TSC4 gene, formerly; TSC4, formerly; Tuberin; Tuberous sclerosis 2; Tuberous sclerosis 2 protein; Tuberous sclerosis 2 protein homolog;

Immunogens

Immunogen:
Uniprot:

P49815(TSC2_HUMAN) >>Visit HPA database.

Gene(ID):
TSC2(7249)
Expression:
P49815 TSC2_HUMAN:

Liver, brain, heart, lymphocytes, fibroblasts, biliary epithelium, pancreas, skeletal muscle, kidney, lung and placenta.

Description:
Mutations in this gene lead to tuberous sclerosis complex. Its gene product is believed to be a tumor suppressor and is able to stimulate specific GTPases. The protein associates with hamartin in a cytosolic complex, possibly acting as a chaperone for hamartin.
Sequence:
MAKPTSKDSGLKEKFKILLGLGTPRPNPRSAEGKQTEFIITAEILRELSMECGLNNRIRMIGQICEVAKTKKFEEHAVEALWKAVADLLQPERPLEARHAVLALLKAIVQGQGERLGVLRALFFKVIKDYPSNEDLHERLEVFKALTDNGRHITYLEEELADFVLQWMDVGLSSEFLLVLVNLVKFNSCYLDEYIARMVQMICLLCVRTASSVDIEVSLQVLDAVVCYNCLPAESLPLFIVTLCRTINVKELCEPCWKLMRNLLGTHLGHSAIYNMCHLMEDRAYMEDAPLLRGAVFFVGMALWGAHRLYSLRNSPTSVLPSFYQAMACPNEVVSYEIVLSITRLIKKYRKELQVVAWDILLNIIERLLQQLQTLDSPELRTIVHDLLTTVEELCDQNEFHGSQERYFELVERCADQRPESSLLNLISYRAQSIHPAKDGWIQNLQALMERFFRSESRGAVRIKVLDVLSFVLLINRQFYEEELINSVVISQLSHIPEDKDHQVRKLATQLLVDLAEGCHTHHFNSLLDIIEKVMARSLSPPPELEERDVAAYSASLEDVKTAVLGLLVILQTKLYTLPASHATRVYEMLVSHIQLHYKHSYTLPIASSIRLQAFDFLLLLRADSLHRLGLPNKDGVVRFSPYCVCDYMEPERGSEKKTSGPLSPPTGPPGPAPAGPAVRLGSVPYSLLFRVLLQCLKQESDWKVLKLVLGRLPESLRYKVLIFTSPCSVDQLCSALCSMLSGPKTLERLRGAPEGFSRTDLHLAVVPVLTALISYHNYLDKTKQREMVYCLEQGLIHRCASQCVVALSICSVEMPDIIIKALPVLVVKLTHISATASMAVPLLEFLSTLARLPHLYRNFAAEQYASVFAISLPYTNPSKFNQYIVCLAHHVIAMWFIRCRLPFRKDFVPFITKGLRSNVLLSFDDTPEKDSFRARSTSLNERPKSLRIARPPKQGLNNSPPVKEFKESSAAEAFRCRSISVSEHVVRSRIQTSLTSASLGSADENSVAQADDSLKNLHLELTETCLDMMARYVFSNFTAVPKRSPVGEFLLAGGRTKTWLVGNKLVTVTTSVGTGTRSLLGLDSGELQSGPESSSSPGVHVRQTKEAPAKLESQAGQQVSRGARDRVRSMSGGHGLRVGALDVPASQFLGSATSPGPRTAPAAKPEKASAGTRVPVQEKTNLAAYVPLLTQGWAEILVRRPTGNTSWLMSLENPLSPFSSDINNMPLQELSNALMAAERFKEHRDTALYKSLSVPAASTAKPPPLPRSNTVASFSSLYQSSCQGQLHRSVSWADSAVVMEEGSPGEVPVLVEPPGLEDVEAALGMDRRTDAYSRSSSVSSQEEKSLHAEELVGRGIPIERVVSSEGGRPSVDLSFQPSQPLSKSSSSPELQTLQDILGDPGDKADVGRLSPEVKARSQSGTLDGESAAWSASGEDSRGQPEGPLPSSSPRSPSGLRPRGYTISDSAPSRRGKRVERDALKSRATASNAEKVPGINPSFVFLQLYHSPFFGDESNKPILLPNESQSFERSVQLLDQIPSYDTHKIAVLYVGEGQSNSELAILSNEHGSYRYTEFLTGLGRLIELKDCQPDKVYLGGLDVCGEDGQFTYCWHDDIMQAVFHIATLMPTKDVDKHRCDKKRHLGNDFVSIVYNDSGEDFKLGTIKGQFNFVHVIVTPLDYECNLVSLQCRKDMEGLVDTSVAKIVSDRNLPFVARQMALHANMASQVHHSRSNPTDIYPSKWIARLRHIKRLRQRICEEAAYSNPSLPLVHPPSHSKAPAQTPAEPTPGYEVGQRKRLISSVEDFTEFV

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Zebrafish
83
Dog
0
Xenopus
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P49815 As Substrate

Site PTM Type Enzyme
K3 Ubiquitination
K7 Acetylation
K7 Ubiquitination
K12 Acetylation
K16 Ubiquitination
T23 Phosphorylation
K34 Ubiquitination
K69 Acetylation
K69 Ubiquitination
K72 Ubiquitination
K106 Ubiquitination
K128 Ubiquitination
K144 Ubiquitination
S188 Phosphorylation
K250 Ubiquitination
K258 Ubiquitination
K438 Ubiquitination
S540 Phosphorylation
K634 Ubiquitination
K658 Ubiquitination
S664 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
K704 Ubiquitination
R751 Methylation
T760 Phosphorylation
K914 Ubiquitination
S923 Phosphorylation
T927 Phosphorylation
K930 Ubiquitination
S932 Phosphorylation
S937 Phosphorylation
T938 Phosphorylation
S939 Phosphorylation P31749 (AKT1) , P31751 (AKT2)
S946 Phosphorylation
K954 Ubiquitination
S960 Phosphorylation
K964 Ubiquitination
K967 Methylation
S981 Phosphorylation P31749 (AKT1)
S999 Phosphorylation
K1043 Ubiquitination
K1058 Ubiquitination
S1096 Phosphorylation
S1097 Phosphorylation
K1106 Ubiquitination
K1111 Ubiquitination
R1122 Methylation
S1130 Phosphorylation
S1132 Phosphorylation
S1155 Phosphorylation
K1168 Ubiquitination
K1180 Ubiquitination
Y1186 Phosphorylation
T1203 Phosphorylation Q13464 (ROCK1)
K1242 Ubiquitination
Y1250 Phosphorylation
K1251 Ubiquitination
S1252 Phosphorylation
S1254 Phosphorylation P49137 (MAPKAPK2)
K1262 Ubiquitination
S1269 Phosphorylation
T1271 Phosphorylation
S1304 Phosphorylation
S1334 Phosphorylation
S1337 Phosphorylation
S1338 Phosphorylation
S1340 Phosphorylation
S1341 Phosphorylation
S1346 Phosphorylation
R1355 Methylation
S1364 Phosphorylation
S1365 Phosphorylation
S1371 Phosphorylation
S1375 Phosphorylation
S1379 Phosphorylation
S1383 Phosphorylation
S1385 Phosphorylation
S1386 Phosphorylation
S1387 Phosphorylation P54646 (PRKAA2)
S1388 Phosphorylation
T1393 Phosphorylation
S1411 Phosphorylation
S1418 Phosphorylation
S1420 Phosphorylation
T1422 Phosphorylation
S1427 Phosphorylation
S1448 Phosphorylation
S1449 Phosphorylation
S1452 Phosphorylation
S1454 Phosphorylation
Y1461 Phosphorylation
T1462 Phosphorylation P31749 (AKT1) , P31751 (AKT2)
S1466 Phosphorylation
S1469 Phosphorylation
Y1571 Phosphorylation
K1585 Ubiquitination
S1647 Phosphorylation
Y1650 Phosphorylation
S1653 Phosphorylation
K1658 Ubiquitination
S1684 Phosphorylation
K1689 Ubiquitination
T1697 Phosphorylation
K1701 Ubiquitination
S1704 Phosphorylation
S1730 Phosphorylation
Y1736 Phosphorylation
K1739 Ubiquitination
Y1760 Phosphorylation
S1761 Phosphorylation
S1764 Phosphorylation
K1775 Ubiquitination
T1780 Phosphorylation
Y1788 Phosphorylation
S1798 Phosphorylation Q15418 (RPS6KA1) , P51812 (RPS6KA3) , Q9P1W9 (PIM2)
S1799 Phosphorylation

Research Backgrounds

Function:

In complex with TSC1, this tumor suppressor inhibits the nutrient-mediated or growth factor-stimulated phosphorylation of S6K1 and EIF4EBP1 by negatively regulating mTORC1 signaling. Acts as a GTPase-activating protein (GAP) for the small GTPase RHEB, a direct activator of the protein kinase activity of mTORC1. May also play a role in microtubule-mediated protein transport (By similarity). Also stimulates the intrinsic GTPase activity of the Ras-related proteins RAP1A and RAB5 (By similarity).

PTMs:

Phosphorylation at Ser-1387, Ser-1418 or Ser-1420 does not affect interaction with TSC1. Phosphorylation at Ser-939 and Thr-1462 by PKB/AKT1 is induced by growth factor stimulation. Phosphorylation by AMPK activates it and leads to negatively regulates the mTORC1 complex. Phosphorylated at Ser-1798 by RPS6KA1; phosphorylation inhibits TSC2 ability to suppress mTORC1 signaling. Phosphorylated by DAPK1.

Ubiquitinated by the DCX(FBXW5) E3 ubiquitin-protein ligase complex, leading to its subsequent degradation. Ubiquitinated by MYCBP2 independently of its phosphorylation status leading to subsequent degradation; association with TSC1 protects from ubiquitination.

Subcellular Location:

Cytoplasm. Membrane>Peripheral membrane protein.
Note: At steady state found in association with membranes.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Liver, brain, heart, lymphocytes, fibroblasts, biliary epithelium, pancreas, skeletal muscle, kidney, lung and placenta.

Subunit Structure:

Probably forms a complex composed of chaperones HSP90 and HSP70, co-chaperones STIP1/HOP, CDC37, PPP5C, PTGES3/p23, TSC1 and client protein TSC2. Probably forms a complex composed of chaperones HSP90 and HSP70, co-chaperones CDC37, PPP5C, TSC1 and client protein TSC2, CDK4, AKT, RAF1 and NR3C1; this complex does not contain co-chaperones STIP1/HOP and PTGES3/p23. Forms a complex containing HSP90AA1, TSC1 and TSC2; TSC1 is required to recruit TCS2 to the complex thereby stabilizing TSC2. Interacts with TSC1 and HERC1; the interaction with TSC1 stabilizes TSC2 and prevents the interaction with HERC1. May also interact with the adapter molecule RABEP1. The final complex may contain TSC2 and RABEP1 linked to RAB5. Interacts with HSPA1 and HSPA8. Interacts with DAPK1. Interacts with FBXW5. Interacts with NAA10 (via C-terminal domain). Interacts with RRAGA (polyubiquitinated). Interacts with WDR45B. Interacts with RPAP3 and URI1.

(Microbial infection) Interacts with human cytomegalovirus protein UL38; this interaction inhibits cellular stress response mediated by mTORC1.

Research Fields

· Cellular Processes > Cell growth and death > p53 signaling pathway.   (View pathway)

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Choline metabolism in cancer.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway.   (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

References

1). Resistance to anti-HER2 therapy associated with the TSC2 nonsynonymous variant c.4349 C > G (p.Pro1450Arg) is reversed by CDK4/6 inhibitor in HER2-positive breast cancer. npj Breast Cancer, 2023 (PubMed: 37160904) [IF=5.9]

Application: WB    Species: Human    Sample: BT474 cell

Fig. 3 Expression of proteins involved in the HER2/AKT/mTOR and cyclinD1/CDK4 signaling pathways in BT474 cell lines treated with lapatinib and/or palbociclib. a Western blot analysis of proteins involved in the HER2/AKT/mTOR signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. b Western blot analysis of proteins involved in the cyclinD1/CDK4 signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. c p-HER2 expression levels under treatment with different drugs in the same cell. d p-HER2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. e p-TSC2 expression levels under treatment with different drugs in the same cell. f p-TSC2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. g p-mTOR expression levels under treatment with different drugs in the same cell. h p-mTOR expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. i p-P70S6K expression levels under treatment with different drugs in the same cell. j p-P70S6K expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. Data are shown as mean ± SD; One-way ANOVA was used to analyze the data in (d, f, h, j). Error bars are SEM.

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