APPL1 Polyclonal antibody

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  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.

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  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.

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  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.

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  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.

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  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).

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  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).

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

Cat No. 12639-1-AP

Publications(10)

Reviews (3)

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

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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

• 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
• 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. 20ul sizes contain 0.1% BSA.

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].

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
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
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
mouse	WB	J Agric Food Chem; Naringin Promotes Skeletal Muscle Fiber Remodeling by the AdipoR1-APPL1-AMPK Signaling Pathway.; Authors - Peiyuan Li
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
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

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Ying (Verified Customer) (09-11-2023)

used for WB and IFC of brain section of wild type and APPL1 transgenic mice

• Applications: Western Blot, Immunohistochemistry, Immunofluorescence, Immunoprecipitation
• Primary Antibody Dilution: 1:500 for IFC, 1:1000 for WB
• Cell Tissue Type: mouse brain/human fibroblast

APPL1 Antibody Western Blot,Immunohistochemistry,Immunofluorescence,Immunoprecipitation validation (1:500 for IFC, 1:1000 for WB dilution) in mouse brain/human fibroblast (Cat no:12639-1-AP)

FH

Sophie (Verified Customer) (11-11-2022)

I used the antibody in a western blot, my samples were primary B-cell lysates (10ug/lane). The antibody did label APPL1 around the 80kDa mark, but I did see another band lower down, around 37kDa - maybe this is a cleavage product?

• Applications: Western Blot
• Primary Antibody Dilution: 1:2,000
• Cell Tissue Type: Primary B-cells (human, from blood)

APPL1 Antibody Western Blot validation (1:2,000 dilution) in Primary B-cells (human, from blood) (Cat no:12639-1-AP)

FH

X (Verified Customer) (01-18-2021)

Very strong and specific signal in WB with mouse brain lysate

• Applications: Western Blot
• Primary Antibody Dilution: 1:5000
• Cell Tissue Type: mouse brain

APPL1 Antibody Western Blot validation (1:5000 dilution) in mouse brain (Cat no:12639-1-AP)