Validation Data Gallery

Filter: AllWBIPIHCIF/ICCFC (Intra)

WB analysis of human brain using 19885-1-AP
human brain tissue were subjected to SDS PAGE followed by western blot with 19885-1-AP (APPL1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of HEK-293 using 19885-1-AP
WB result of APPL1 antibody (19885-1-AP; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-APPL1 transfected HEK-293 cells.
WB analysis of HT-1080 using 19885-1-AP
HT-1080 cells were subjected to SDS PAGE followed by western blot with 19885-1-AP (APPL1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.
WB analysis of HeLa using 19885-1-AP
HeLa cells were subjected to SDS PAGE followed by western blot with 19885-1-AP (APPL1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of human heart using 19885-1-AP
human heart tissue were subjected to SDS PAGE followed by western blot with 19885-1-AP (APPL1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.
WB analysis of mouse brain using 19885-1-AP
mouse brain tissue were subjected to SDS PAGE followed by western blot with 19885-1-AP (APPL1 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of HEK-293 using 19885-1-AP
HEK-293 cells were subjected to SDS PAGE followed by western blot with 19885-1-AP (APPL1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
IP experiment of mouse brain using 19885-1-AP
IP result of anti-APPL1 (IP:19885-1-AP, 4ug; Detection:19885-1-AP 1:1000) with mouse brain tissue lysate 2640ug.
IHC staining of human breast cancer using 19885-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer slide using 19885-1-AP (APPL1 Antibody) at dilution of 1:50.
IF Staining of HepG2 using 19885-1-AP
Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using APPL1 antibody (19885-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).
IF Staining of HepG2 using 19885-1-AP
Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using APPL1 antibody (19885-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).
FC experiment of HepG2 using 19885-1-AP
1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human APPL1 (19885-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

Featured Product
KD/KO Validated

APPL1 Polyclonal antibody

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

Cat No. 19885-1-AP

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, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

CoraLite® Plus 488

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	human brain tissue, HEK-293 cells, HeLa cells, human heart tissue, HT-1080 cells, mouse brain tissue
Positive IP detected in	mouse brain tissue
Positive IHC detected in	human breast cancer 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 : 1:1000-1:4000
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.

Product Information

19885-1-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
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag13703 Product name: Recombinant human APPL1 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-323 aa of BC028599 Sequence: MPGIDKLPIEETLEDSPQTRSLLGVFEEDATAISNYMNQLYQAMHRIYDAQNELSAATHLTSKLLKEYEKQRFPLGGDDEVMSSTLQQFSKVIDELSSCHAVLSTQLADAMMFPITQFKERDLKEILTLKEVFQIASNDHDAAINRYSRLSKKRENDKVKYEVTEDVYTSRKKQHQTMMHYFCALNTLQYKKKIALLEPLLGYMQAQISFFKMGSENLNEQLEEFLANIGTSVQNVRREMDSDIETMQQTIEDLEVASDPLYVPDPDPTKFPVNRNLTRKAGYLNARNKTGLVSSTWDRQFYFTQGGNLMSQARGDVAGGLAM 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_10637848
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q9UKG1
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}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
FC protocol for APPL1 antibody 19885-1-AP	Download protocol
IF protocol for APPL1 antibody 19885-1-AP	Download protocol
IHC protocol for APPL1 antibody 19885-1-AP	Download protocol
IP protocol for APPL1 antibody 19885-1-AP	Download protocol
WB protocol for APPL1 antibody 19885-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis of human brain using 19885-1-AP

WB analysis of HEK-293 using 19885-1-AP

WB analysis of HT-1080 using 19885-1-AP

WB analysis of HeLa using 19885-1-AP

WB analysis of human heart using 19885-1-AP

WB analysis of mouse brain using 19885-1-AP