Validation Data Gallery

Filter: AllWBIPIHCIF/ICCFC (Intra)

WB analysis using 12639-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 12639-1-AP (APPL1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.
WB analysis of HEK-293 using 12639-1-AP
WB result of APPL1 antibody (12639-1-AP; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-APPL1 transfected HEK-293 cells.
WB analysis of mouse ovary using 12639-1-AP
mouse ovary tissue were subjected to SDS PAGE followed by western blot with 12639-1-AP (APPL1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.
IP experiment of mouse brain using 12639-1-AP
IP result of anti-APPL1 (IP:12639-1-AP, 4ug; Detection:12639-1-AP 1:500) with mouse brain tissue lysate 4000ug.
IHC staining of human ovary tumor using 12639-1-AP
Immunohistochemical analysis of paraffin-embedded human ovary tumor using 12639-1-AP (APPL1 antibody) at dilution of 1:50 (under 10x lens).
IF Staining of HepG2 using 12639-1-AP
Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using APPL1 antibody (12639-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).
FC experiment of HepG2 using 12639-1-AP
1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human APPL1 (12639-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Featured Product
KD/KO Validated

APPL1 Polyclonal antibody

APPL1 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 12639-1-AP

Publications(10)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Synonyms

DIP13alpha, DIP13A, Dip13 alpha, DCC-interacting protein 13-alpha, APPL

Formulation: PBS and Azide

PBS and Azide

Conjugate: Unconjugated

Unconjugated

CoraLite® Plus 488

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (10)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	C2C12 cells, HEK-293 cells, mouse ovary tissue, HeLa cells, mouse liver tissue
Positive IP detected in	mouse brain tissue
Positive IHC detected in	human ovary tumor tissue **Note: suggested antigen retrieval with TE buffer pH 9.0; () Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0***
Positive IF/ICC detected in	HepG2 cells
Positive FC (Intra) detected in	HepG2 cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:2000-1:10000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)	IHC : 1:20-1:200
Immunofluorescence (IF)/ICC	IF/ICC : 1:50-1:500
Flow Cytometry (FC) (INTRA)	FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Published Applications

KD/KO	See 1 publications below
WB	See 9 publications below
IHC	See 1 publications below
IF	See 3 publications below

Product Information

12639-1-AP targets APPL1 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human, mouse, rat
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	APPL1 fusion protein Ag3334 相同性解析による交差性が予測される生物種
Full Name	adaptor protein, phosphotyrosine interaction, PH domain and leucine zipper containing 1
Calculated molecular weight	709 aa, 80 kDa
Observed molecular weight	80 kDa
GenBank accession number	BC028599
Gene Symbol	APPL1
Gene ID (NCBI)	26060
RRID	AB_2289669
Conjugate	Unconjugated
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q9UKG1
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol , pH 7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

Adaptor protein, phosphotyrosine interaction, PH domain and leucine zipper containing 1 (APPL1), a binding partner of Akt2 and an important regulator of INS signaling, plays a key role in the regulation of INS secretion [PMID:22615370]. APPL1 interacts with adiponectin receptors and mediates the INS-sensitizing effects of adiponectin in muscle and endothelial cells. It also participates in nuclear signaling and transcriptional regulation, mostly by modulating the activity of various nuclear factors [PMID:22685329]. Apart from its role in endocytosis and endosomal transport, APPL1 was reported to undergo nucleocytoplasmic shuttling and participate in transcriptional regulation, e.g. by interactions with histone deacetylases (HDACs) [PMID:19686092].

Protocols

Product Specific Protocols
WB protocol for APPL1 antibody 12639-1-AP	Download protocol
IHC protocol for APPL1 antibody 12639-1-AP	Download protocol
IF protocol for APPL1 antibody 12639-1-AP	Download protocol
IP protocol for APPL1 antibody 12639-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
rat	WB	Oxid Med Cell Longev DDAH1/ADMA Regulates Adiponectin Resistance in Cerebral Ischemia via the ROS/FOXO1/APR1 Pathway. Authors - Yichen Zhao View Article
human	IF	J Cell Sci ESCRT-0 marks an APPL1-independent transit route for EGFR between the cell surface and the EEA1-positive early endosome. Authors - Neftali Flores-Rodriguez View Article
human	WB	J Biol Chem Endocytosis is required for C-X-C chemokine receptor type 4 (CXCR4)-mediated Akt activation and anti-apoptotic signaling. Authors - Elizabeth J English KD Validated View Article
mouse	WB	J Agric Food Chem Naringin Promotes Skeletal Muscle Fiber Remodeling by the AdipoR1-APPL1-AMPK Signaling Pathway. Authors - Peiyuan Li View Article
mouse	WB,IF	Biol Reprod Myosin VI maintains the actin-dependent organization of the tubulobulbar complexes required for endocytosis during mouse spermiogenesis. Authors - Przemysław Zakrzewski View Article
human	WB,IHC	Brain Res Characterization and distribution of adaptor protein containing a PH domain, PTB domain and leucine zipper motif (APPL1) in Alzheimer's disease hippocampus: an immunohistochemical study. Authors - Akari Ogawa View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 12639-1-AP

WB analysis of HEK-293 using 12639-1-AP

WB analysis of mouse ovary using 12639-1-AP

IP experiment of mouse brain using 12639-1-AP

IHC staining of human ovary tumor using 12639-1-AP

IF Staining of HepG2 using 12639-1-AP

FC experiment of HepG2 using 12639-1-AP

APPL1 Polyclonal antibody

APPL1 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

DDAH1/ADMA Regulates Adiponectin Resistance in Cerebral Ischemia via the ROS/FOXO1/APR1 Pathway.

ESCRT-0 marks an APPL1-independent transit route for EGFR between the cell surface and the EEA1-positive early endosome.

Endocytosis is required for C-X-C chemokine receptor type 4 (CXCR4)-mediated Akt activation and anti-apoptotic signaling.

Naringin Promotes Skeletal Muscle Fiber Remodeling by the AdipoR1-APPL1-AMPK Signaling Pathway.

Myosin VI maintains the actin-dependent organization of the tubulobulbar complexes required for endocytosis during mouse spermiogenesis.

Characterization and distribution of adaptor protein containing a PH domain, PTB domain and leucine zipper motif (APPL1) in Alzheimer's disease hippocampus: an immunohistochemical study.