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APPL1 Monoclonal antibody

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

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat, pig, rabbit

Applications

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

Conjugate

Unconjugated

CloneNo.

1B7B11

Cat no : 68195-1-Ig

Synonyms

DIP13A, Dip13 alpha, DCC-interacting protein 13-alpha, APPL, Adapter protein containing PH domain, PTB domain and leucine zipper motif 1

Validation Data Gallery

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  • Western Blot (WB) analysis of various lysates using APPL1 Monoclonal antibody (68195-1-Ig)

    WB analysis using 68195-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 68195-1-Ig (APPL1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

  • Western Blot (WB) analysis of various lysates using APPL1 Monoclonal antibody (68195-1-Ig)

    WB analysis using 68195-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 68195-1-Ig (APPL1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

  • Western Blot (WB) analysis of various lysates using APPL1 Monoclonal antibody (68195-1-Ig)

    WB analysis using 68195-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 68195-1-Ig (APPL1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

  • Western Blot (WB) analysis of HepG2 cells using APPL1 Monoclonal antibody (68195-1-Ig)

    WB analysis of HepG2 using 68195-1-Ig

    HepG2 cells were subjected to SDS PAGE followed by western blot with 68195-1-Ig (APPL1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

  • Western Blot (WB) analysis of HEK-293 cells using APPL1 Monoclonal antibody (68195-1-Ig)

    WB analysis of HEK-293 using 68195-1-Ig

    WB result of APPL1 antibody (68195-1-Ig; 1:30000; incubated at room temperature for 1.5 hours) with sh-Control and sh-APPL1 transfected HEK-293 cells.

  • Western Blot (WB) analysis of pig heart tissue using APPL1 Monoclonal antibody (68195-1-Ig)

    WB analysis of pig heart using 68195-1-Ig

    pig heart tissue were subjected to SDS PAGE followed by western blot with 68195-1-Ig (APPL1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

  • Immunohistochemistry (IHC) staining of human colon cancer tissue using APPL1 Monoclonal antibody (68195-1-Ig)

    IHC staining of human colon cancer using 68195-1-Ig

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 68195-1-Ig (APPL1 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunofluorescence (IF) / fluorescent staining of HepG2 cells using APPL1 Monoclonal antibody (68195-1-Ig)

    IF Staining of HepG2 using 68195-1-Ig

    Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using APPL1 antibody (68195-1-Ig, Clone: 1B7B11 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

  • Flow cytometry (FC) experiment of HepG2 cells using APPL1 Monoclonal antibody (68195-1-Ig)

    FC experiment of HepG2 using 68195-1-Ig

    1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human APPL1 (68195-1-Ig, Clone:1B7B11) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b Isotype Control (MPC-11) (65128-1-Ig, Clone: MPC-11) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Tested Applications

Positive WB detected inLNCaP cells, Jurkat cells, rabbit heart tissue, HepG2 cells, pig heart tissue, HEK-293 cells, HeLa cells, rat heart tissue
Positive IHC detected inhuman colon 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 inHepG2 cells
Positive FC (Intra) detected inHepG2 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunohistochemistry (IHC)IHC : 1:200-1:800
Immunofluorescence (IF)/ICCIF/ICC : 1:200-1:800
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

68195-1-Ig targets APPL1 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat, pig, rabbit samples.

Tested Reactivity human, mouse, rat, pig, rabbit
Host / Isotype Mouse / IgG2b
Class Monoclonal
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 Weight80 kDa
GenBank Accession NumberBC028599
Gene Symbol APPL1
Gene ID (NCBI) 26060
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC 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].

Protocols

Product Specific Protocols
WB protocol for APPL1 antibody 68195-1-IgDownload protocol
IHC protocol for APPL1 antibody 68195-1-IgDownload protocol
IF protocol for APPL1 antibody 68195-1-IgDownload protocol
FC protocol for APPL1 antibody 68195-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols