anti-phospho-p62 (sqstm1) (ser351) pab phospho...
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Anti-Phospho-p62 (SQSTM1) (Ser403) mAb
Anti-Phospho-p62 (SQSTM1) (Ser351) pAb
Lane 1: GFP-tagged human p62/Neuro2a Lane 2: GFP-tagged human p62/Neuro2a, Bafilomycin A1-treated (1 μM, 24 hr.)Lane 3: MEFAtg5-/- Lane 4: MEF
Lane 1: MEF, sodium arsenite-treated (10 μM, 12 hr.)Lane 2: MEF Lane 3: MEFAtg5-/-
Lane 4: huH-1 Lane 5: huH-1, λ-phosphatase-treatedLane 6: p62-knockout huH-1
Brown: Anti-Phospho-p62 (SQSTM1) (Ser403) mAb (D344-3)Blue: Hematoxylin
Green: Anti-Phospho-p62 (SQSTM1) (Ser351) pAb (PM074)Blue: DAPI
Western blotting（Code No. D343-3）1 3 42
GFP-tagged Phospho-p62
Phospho-p62
Atg5 conditional knockout mouse brain Wild type
Immunohistochemistry（Code No. D344-3）
Western blotting（Code No. PM074） Immunocytochemistry（Code No. PM074）
(kDa)15010075
50
25
37
20
(kDa)150
10075
50
37
1 3 4 5 62
Phospho-p62(Ser351)
p62 TBzzPB1 UBALIR KIR
PP
LC3
p62
p62p62
p62
Pp62
UbUb
UbUb P
p62Ub
UbUbUb
p62
CK2phosphatase
p62 P
P
S403
p62
UbUb
UbUb Ub
Ub
UbUb
Ub
UbUbUb Ub
Ub
UbU
bU
bUb
UbUbUb
Pp62
UbUb
UbUb
Pp62UbUbUbUb
Pp62UbUbUbUb
Pp62
Ub UbUb
UbP
p62Ub Ub
UbUb
Ub Ub
UbUbUbUb
UbUbUbUb
UbUb
UbUb×
Pp62
UbUb
UbUb
Pp62
UbUb
UbUb
PP
P p62
p62
p62
p62
Pp62
UbUb
UbUb
UbUb
UbUb
p62
Pp62
UbUb
UbUb
LC3LC
3
LC3 LC3LC3 LC3
LC3LC3
LC3
LC3LC3
LC3
p62LC3
P
PP
Keap1Nrf2
p62
CK2TBK1
p62P
LC3
LC3
U
U
U
U
UU
Ub
Ub
Ub
p62PP
p62PP
p62PP
Keap1Keap1
Keap1
Nrf2 Nrf2
Nrf2
Maf Nrf2
UU
U
U
U
UUb
Ub
UbU
U
U
UUb
Ub
Ub
p62P
p62P
p62P
U
U
U
UUb
Ub
Ub
p62P
p62P
p62PP
P P
Nrf2
U
U
U
UUb
Ub
Ub
p62P
p62P
p62PP
P P
Keap1Keap1
Keap1
p62p62p62
p62p62
Nrf2
Maf Nrf2
p62 p62
p62
p62
Nrf2
Keap1Nrf2
Nrf2
p62p62p62
p62p62
P
P
P
P
P
Keap1 Keap1
Keap1Keap1
p62p62p62
p62p62
P
P
P
P
P
栄養
源の
供給
を介
した増
殖促
進
NonphosphorylatedS403
Phosphprylated S403
p62 strongly binds to ubiquitin chains
Sequestosome(Isolation of ubiquitinated proteins)
autophagosome
Abnormal proteins that cannot be degraded by proteasomes
Normal cells
Cancer cells
mTORC1Other kinase
Ubiquitinated proteinCytoplasm
Nucleus
Cytoplasm
Nucleus
Nrf2 target genes
Nrf2 target genes
Expression of Nrf2 target genes mediates host defence
Expression of Nrf2 target genes mediates promotion of proliferation
Kinase
Self-assembling
Autophagosome
Amino acidLipid
Carbohydrate
Isolation membrane / phagophore
For Research Use Only
http://ruo.mbl.co.jp/
Code No. Product name Clone Isotype Size Applications Reactivity
D343-3 Anti-Phospho-p62 (SQSTM1) (Ser403) mAb 4F6 Rat IgG2aκ 100 μg WB / IH Hu / Mo
D344-3 Anti-Phospho-p62 (SQSTM1) (Ser403) mAb 4C8 Rat IgG2aκ 100 μg WB / IH Hu / Mo
Phospho-p62 antibodies
p62/SQSTM1 binds directly with LC3 and is selectively degraded by autophagy.It has been proposed that p62 binds to ubiquitinated protein aggregates and it works as an adapter mole-cule that leads the aggregates to autophagosomes.Recently, p62 has been found to be phosphorylated at various residues, and the phosphorylation at each residue regulates different aspects of p62 functions.
Ser351 (mouse) Ser405 (mouse)Ser349 (human) Ser403 (human)
It was reported that when p62 is phosphorylated at Ser403, the affinity between the polyubiquitin chain and p62 increases, thus promoting degradation of the polyubiquitinated proteins through autophagy.
Neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease, are considered to be caused by impaired cellular function resulting from abnormal accumulation of poly-ubiquitinalted proteins (aggregates) in cells.
These findings suggest the possilibity that the development of drugs that specifically promote phosphorylation of Ser403 in p62 may lead to the treatment of the neurodegenerative diseases
The figure was provided by Prof. Nobuyuki Nukina and Dr. Gen Matsumoto (Juntendo University) and partially modified with their permission.
Anti-Phospho-p62 (SQSTM1) (Ser403) mAb
Anti-Phospho-p62 (SQSTM1) (Ser351) pAb
Lane 1: GFP-tagged human p62/Neuro2a Lane 2: GFP-tagged human p62/Neuro2a, Bafilomycin A1-treated (1 μM, 24 hr.)Lane 3: MEFAtg5-/- Lane 4: MEF
Lane 1: MEF, sodium arsenite-treated (10 μM, 12 hr.)Lane 2: MEF Lane 3: MEFAtg5-/-
Lane 4: huH-1 Lane 5: huH-1, λ-phosphatase-treatedLane 6: p62-knockout huH-1
Brown: Anti-Phospho-p62 (SQSTM1) (Ser403) mAb (D344-3)Blue: Hematoxylin
Green: Anti-Phospho-p62 (SQSTM1) (Ser351) pAb (PM074)Blue: DAPI
Western blotting（Code No. D343-3）1 3 42
GFP-tagged Phospho-p62
Phospho-p62
Atg5 conditional knockout mouse brain Wild type
Immunohistochemistry（Code No. D344-3）
Western blotting（Code No. PM074） Immunocytochemistry（Code No. PM074）
(kDa)150100
75
50
25
37
20
(kDa)150
10075
50
37
1 3 4 5 62
Phospho-p62(Ser351)
p62 TBzzPB1 UBALIR KIR
PP
LC3
p62
p62p62
p62
Pp62
UbUb
UbUb P
p62Ub
UbUbUb
p62
CK2phosphatase
p62 P
P
S403
p62
UbUb
UbUb Ub
Ub
UbUb
Ub
UbUbUb Ub
Ub
UbU
bU
b
UbUb
UbUb
Pp62
UbUb
UbUb
Pp62UbUbUbUb
Pp62UbUbUbUb
Pp62
Ub UbUb
UbP
p62Ub Ub
UbUb
Ub Ub
UbUbUbUb
UbUbUbUb
UbUb
UbUb×
Pp62
UbUb
UbUb
Pp62
UbUb
UbUb
PP
P p62
p62
p62
p62
Pp62
UbUb
UbUb
UbUb
UbUb
p62
Pp62
UbUb
UbUb
LC3LC
3
LC3 LC3LC3 LC3
LC3LC3
LC3
LC3LC3
LC3
p62LC3
P
PP
Keap1Nrf2
p62
CK2TBK1
p62P
LC3
LC3
U
U
U
U
UU
Ub
Ub
Ub
p62PP
p62PP
p62PP
Keap1Keap1
Keap1
Nrf2 Nrf2
Nrf2
Maf Nrf2
UU
U
U
U
UUb
Ub
UbU
U
U
UUb
Ub
Ub
p62P
p62P
p62P
U
U
U
UUb
Ub
Ub
p62P
p62P
p62PP
P P
Nrf2
U
U
U
UUb
Ub
Ub
p62P
p62P
p62PP
P P
Keap1Keap1
Keap1
p62p62p62
p62p62
Nrf2
Maf Nrf2
p62 p62
p62
p62
Nrf2
Keap1Nrf2
Nrf2
p62p62p62
p62p62
P
P
P
P
P
Keap1 Keap1
Keap1Keap1
p62p62p62
p62p62
P
P
P
P
P
栄養
源の
供給
を介
した増
殖促
進
NonphosphorylatedS403
Phosphprylated S403
p62 strongly binds to ubiquitin chains
Sequestosome(Isolation of ubiquitinated proteins)
autophagosome
Abnormal proteins that cannot be degraded by proteasomes
Normal cells
Cancer cells
mTORC1Other kinase
Ubiquitinated proteinCytoplasm
Nucleus
Cytoplasm
Nucleus
Nrf2 target genes
Nrf2 target genes
Expression of Nrf2 target genes mediates host defence
Expression of Nrf2 target genes mediates promotion of proliferation
Kinase
Self-assembling
Autophagosome
Amino acidLipid
Carbohydrate
Isolation membrane / phagophore
146065-140712014.07
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MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
Produced by
Code No. Product name Clone Isotype Size Applications Reactivity
PM074 Anti-Phospho-p62 (SQSTM1) (Ser351) pAb Polyclonal Rabbit Ig (aff.) 100 μL WB / IC / IH Hu / Mo
Related products
Code No. Product name Clone Isotype Size Applications Reactivity
M162-3 Anti-p62 (SQSTM1) (Human) mAb 5F2 Mouse IgG1κ 100 μg WB / IP / FCM / IC / IH Hu
M162-A48 Anti-p62 (SQSTM1) (Human) mAb-Alexa Fluor® 488 5F2 Mouse IgG1κ 100 μg FCM / IC Hu
M162-A59 Anti-p62 (SQSTM1) (Human) mAb-Alexa Fluor® 594 5F2 Mouse IgG1κ 100 μg IC Hu
M162-A64 Anti-p62 (SQSTM1) (Human) mAb-Alexa Fluor® 647 5F2 Mouse IgG1κ 100 μg FCM / IC Hu
PM045 Anti-p62 (SQSTM1) pAb Polyclonal Rabbit Ig(aff.) 100 μL WB / IP / IC / IH Hu / Mo / Rat / Ham
PM066 Anti-p62 C-terminal pAb Polyclonal Guinea pig Ig (aff.) 100 μL WB / IP / IC / IH Hu / Mo / Rat / Ham
PM066-7 Anti-p62 C-terminal pAb-HRP-DirecT Polyclonal Guinea pig Ig (aff.) 50 μL WB Hu / Mo / Rat / Ham
Applications: WB: Western Blotting IP: Immunoprecipitation FCM: Flow Cytometry IC: Immunocytochemistry IH: ImmunohistochemistryReactivity: Hu: Human Mo: Mouse Ham: HamsterAlexa Fluor® is the registered trademark of Life Technologies. MBL has received from Life Technologies the license to produce and market Alexa Fluor®-conjugated antibodies.
The figures were provided by Prof. Masaaki Komatsu (Niigata University) and partially modified with permission from Prof. Komatsu.
Phosphorylation of p62 at Ser351 causes significant increases in binding affinity to Keap1, a ubiquitin ligase adaptor protein.
The binding of phospho-p62 (Ser351) to Keap1 stabilizes a transcription factor Nrf2. Stabilized Nrf2 translocates to the nuc leus and induces the expression of antioxidant genes. (The phospho-p62/Keap1 complex is degraded by autophagy)
Furthermore, Nrf2 stabilization also causes an increase in the expression of cell proliferation-related genes in cancer cells. Therefore, inhibitors of the interaction between phosphorylated p62 (Ser351) and Keap1 may lead to potential therapeutic drugs for treatment of cancers.