Phospho-DEPDC5 (Ser1530) Antibody | Affinity Biosciences

pElisa
Cites

1/2

Product:	Phospho-DEPDC5 (Ser1530) Antibody
Catalog:	AF8617
Description:	Rabbit polyclonal antibody to Phospho-DEPDC5 (Ser1530)
Application:	WB
Reactivity:	Human
Prediction:	Zebrafish, Bovine, Horse, Rabbit, Dog, Chicken
Mol.Wt.:	180 kDa; 181kD(Calculated).
Uniprot:	O75140
RRID:	AB_2844377

View similar products>>

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$350	In stock
200ul	$450	In stock

Lead Time: Same day delivery

For pricing and ordering contact:
Local distributors

Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:1000-3000
*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

Prediction:

Zebrafish(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%)

Clonality:

Polyclonal

Specificity:

Phospho-DEPDC5 (Ser1530) Antibody detects endogenous levels of DEPDC5 only when phosphorylated at S1530.

RRID:

AB_2844377
Cite Format: Affinity Biosciences Cat# AF8617, RRID:AB_2844377.

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

DEP domain containing 5; DEP domain-containing protein 5; DEP.5; DEPD5_HUMAN; DEPDC5; FFEVF; KIAA0645;

Immunogens

Immunogen:

Uniprot:

O75140(DEPD5_HUMAN) >>Visit HPA database.

Gene(ID):

DEPDC5(9681)

Expression:

O75140 DEPD5_HUMAN:

Expressed in developing and adult brain.

Sequence:

O75140 DEPD5_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MRTTKVYKLVIHKKGFGGSDDELVVNPKVFPHIKLGDIVEIAHPNDEYSPLLLQVKSLKEDLQKETISVDQTVTQVFRLRPYQDVYVNVVDPKDVTLDLVELTFKDQYIGRGDMWRLKKSLVSTCAYITQKVEFAGIRAQAGELWVKNEKVMCGYISEDTRVVFRSTSAMVYIFIQMSCEMWDFDIYGDLYFEKAVNGFLADLFTKWKEKNCSHEVTVVLFSRTFYDAKSVDEFPEINRASIRQDHKGRFYEDFYKVVVQNERREEWTSLLVTIKKLFIQYPVLVRLEQAEGFPQGDNSTSAQGNYLEAINLSFNVFDKHYINRNFDRTGQMSVVITPGVGVFEVDRLLMILTKQRMIDNGIGVDLVCMGEQPLHAVPLFKLHNRSAPRDSRLGDDYNIPHWINHSFYTSKSQLFCNSFTPRIKLAGKKPASEKAKNGRDTSLGSPKESENALPIQVDYDAYDAQVFRLPGPSRAQCLTTCRSVRERESHSRKSASSCDVSSSPSLPSRTLPTEEVRSQASDDSSLGKSANILMIPHPHLHQYEVSSSLGYTSTRDVLENMMEPPQRDSSAPGRFHVGSAESMLHVRPGGYTPQRALINPFAPSRMPMKLTSNRRRWMHTFPVGPSGEAIQIHHQTRQNMAELQGSGQRDPTHSSAELLELAYHEAAGRHSNSRQPGDGMSFLNFSGTEELSVGLLSNSGAGMNPRTQNKDSLEDSVSTSPDPILTLSAPPVVPGFCCTVGVDWKSLTTPACLPLTTDYFPDRQGLQNDYTEGCYDLLPEADIDRRDEDGVQMTAQQVFEEFICQRLMQGYQIIVQPKTQKPNPAVPPPLSSSPLYSRGLVSRNRPEEEDQYWLSMGRTFHKVTLKDKMITVTRYLPKYPYESAQIHYTYSLCPSHSDSEFVSCWVEFSHERLEEYKWNYLDQYICSAGSEDFSLIESLKFWRTRFLLLPACVTATKRITEGEAHCDIYGDRPRADEDEWQLLDGFVRFVEGLNRIRRRHRSDRMMRKGTAMKGLQMTGPISTHSLESTAPPVGKKGTSALSALLEMEASQKCLGEQQAAVHGGKSSAQSAESSSVAMTPTYMDSPRKDGAFFMEFVRSPRTASSAFYPQVSVDQTATPMLDGTSLGICTGQSMDRGNSQTFGNSQNIGEQGYSSTNSSDSSSQQLVASSLTSSSTLTEILEAMKHPSTGVQLLSEQKGLSPYCFISAEVVHWLVNHVEGIQTQAMAIDIMQKMLEEQLITHASGEAWRTFIYGFYFYKIVTDKEPDRVAMQQPATTWHTAGVDDFASFQRKWFEVAFVAEELVHSEIPAFLLPWLPSRPASYASRHSSFSRSFGGRSQAAALLAATVPEQRTVTLDVDVNNRTDRLEWCSCYYHGNFSLNAAFEIKLHWMAVTAAVLFEMVQGWHRKATSCGFLLVPVLEGPFALPSYLYGDPLRAQLFIPLNISCLLKEGSEHLFDSFEPETYWDRMHLFQEAIAHRFGFVQDKYSASAFNFPAENKPQYIHVTGTVFLQLPYSKRKFSGQQRRRRNSTSSTNQNMFCEERVGYNWAYNTMLTKTWRSSATGDEKFADRLLKDFTDFCINRDNRLVTFWTSCLEKMHASAP

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

Horse

100

Bovine

100

Dog

100

Zebrafish

100

Chicken

100

Rabbit

100

Pig

Sheep

Xenopus

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O75140 As Substrate

O75140

Site	PTM Type	Source
T66	Phosphorylation	Uniprot
Y155	Phosphorylation	Uniprot
T273	Phosphorylation	Uniprot
K411	Methylation	Uniprot
S432	Phosphorylation	Uniprot
S445	Phosphorylation	Uniprot
S489	Phosphorylation	Uniprot
S491	Phosphorylation	Uniprot
S494	Phosphorylation	Uniprot
S496	Phosphorylation	Uniprot
S497	Phosphorylation	Uniprot
S503	Phosphorylation	Uniprot
S505	Phosphorylation	Uniprot
S508	Phosphorylation	Uniprot
S521	Phosphorylation	Uniprot
S524	Phosphorylation	Uniprot
S525	Phosphorylation	Uniprot
S548	Phosphorylation	Uniprot
S579	Phosphorylation	Uniprot
T749	Phosphorylation	Uniprot
Y759	Phosphorylation	Uniprot
S831	Phosphorylation	Uniprot
S833	Phosphorylation	Uniprot
Y920	Phosphorylation	Uniprot
Y924	Phosphorylation	Uniprot
S1028	Phosphorylation	Uniprot
S1530	Phosphorylation	Uniprot
T1531	Phosphorylation	Uniprot
Y1546	Phosphorylation	Uniprot
Y1550	Phosphorylation	Uniprot
K1574	Ubiquitination	Uniprot

Research Backgrounds

O75140_DEPD5_HUMAN

Function:

As a component of the GATOR1 complex functions as an inhibitor of the amino acid-sensing branch of the TORC1 pathway. The GATOR1 complex strongly increases GTP hydrolysis by RRAGA and RRAGB within RRAGC-containing heterodimers, thereby deactivating RRAGs, releasing mTORC1 from lysosomal surface and inhibiting mTORC1 signaling. The GATOR1 complex is negatively regulated by GATOR2 the other GATOR subcomplex in this amino acid-sensing branch of the TORC1 pathway.

PTMs:

Ubiquitinated. Amino acid-induced 'Lys-48'-linked polyubiquitination of DEPDC5 by the BCR(KLHL22) ubiquitin ligase complex leads to DEPDC5 proteasomal degradation and inhibition of the GATOR1 complex. Ubiquitination may occur at multiple lysines.

Subcellular Location:

Cytoplasm>Cytosol. Cytoplasm>Perinuclear region. Lysosome membrane.
Note: Localization to lysosomes is amino acid-independent.

Tissue Specificity:

Expressed in developing and adult brain.

Subunit Structure:

Within the GATOR complex, component of the GATOR1 subcomplex, made of DEPDC5, NPRL2 and NPRL3. GATOR1 mediates the strong interaction of the GATOR complex with RRAGA/RRAGC and RRAGB/RRAGC heterodimers. Interacts (via DEP domain) with KLHL22; the interaction depends on amino acid availability.

Family&Domains:

The DEP domain mediates the interaction with KLHL22.

Belongs to the IML1 family.

Research Fields

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

References

1). Phosphorylation of DEPDC5, a component of the GATOR1 complex, releases inhibition of mTORC1 and promotes tumor growth. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2019 (PubMed: 31548394) [IF=11.1]

Application: WB Species: Human Sample: 293T cells

Fig. 2. Pim and AKT kinases phosphorylate DEPDC5. (A) Pim1 and Flag-tagged DEPDC5 (Flag DEPDC5) plasmids were cotransfected into 293T cells. Flag immunoprecipitated (IP) samples and whole cell lysates (WCL) were analyzed by WB using the indicated antibodies. (B and C) Plasmids expressing DEPDC5 WT, S1002A (SA1), and S1530A (SA2) were transfected into 293T cells which were treated with and without 3 µM Pim447 for 16 h and then cells were lysed. WCL and Flag-IP samples were analyzed by WB using the indicated antibodies. (D and E) Plasmids expressing Flag DEPDC5, Pim1, or myr-AKT were cotransfected into 293T cells and then treated with 3 µM Pim447 or the AKT inhibitor AZD5363, 3 µM for 18 h. The Flag-IP samples and WCL were analyzed by WB. (F) BT549 cells stably expressing EGFP as a control or DEPDC5 were incubated with DMSO (C), 3 µM Pim447 (P), 3 µM AZD5363 (A), and the combination (AP) for 6 h and cell lysates were analyzed by WB using the indicated antibodies. (G) BT549 cells stably expressing DEPDC5 were starved for 24 h and pretreated with DMSO (C), 3 µM Pim447 (P), 5 µM GSK690693 (A), and the combination (AP) for 1.5 h before stimulation with insulin (0.5 µg/mL) for 30 min, and cell lysates were analyzed by WB using the indicated antibodies. (H) BT549 cells stably expressing DEPDC5 were transfected with siRNA targeting Pim1 or all 3 Pim kinases, and after 48 h, cell lysates were analyzed by WB. See SI Appendix, SI Materials and Methods for WB quantification. Numerical values shown are calculated relative to the DMSO (C) treatment for F or no insulin treatment for G.

Restrictive clause

Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

For Research Use Only.
Not for use in diagnostic or therapeutic procedures. Not for resale. Not for distribution without written consent. Affinity Biosciences will not be held responsible for patent infringement or other violations that may occur with the use of our products. Affinity Biosciences, Affinity Biosciences Logo and all other trademarks are the property of Affinity Biosciences LTD.

Phospho-DEPDC5 (Ser1530) Antibody - #AF8617