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  4. APPL Antibody (YA2135)

APPL Antibody (YA2135) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to APPL.

For research use only. We do not sell to patients.

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Top Publications Citing Use of Products
  • WB: Western Blot;
  • IHC-P: Immunohistochemistry-Paraffin;
  • IHC-F: Immunohistochemistry-Frozen;
  • ICC/IF: Immunocytochemistry/Immunofluorescence;
  • IF-Tissue: Immunofluorescence-Tissue;
  • mIHC: Multiplex Immunohistochemical;
  • IP: Immunoprecipitation;
  • ChIP: Chromatin Immunoprecipitation;
  • FC: Flow Cytometry;
  • ELISA: Enzyme Linked Immunosorbent Assay
  • Product Detail

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Description

APPL Antibody (YA2135) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to APPL.

Host

Rabbit

Clonality

Recombinant,Monoclonal

Molecular Weight
Predicted band size: 80 kDa;
Observed band size: 80 kDa
Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
Species Reactivity
Human, Mouse, Rat
SwissProt ID
Gene ID
Immunogen

A synthetic peptide of human APPL

Application &
Dilution Ratio
Application Dilution Ratio
WB
WB: Western Blot
1:500-1:1000
IHC-P
IHC-P: Immunohistochemistry-Paraffin
1:50-1:100
Sensitivity Endogenous Purity Affinity Purified
Conjugation Non-conjugated Modification Unmodified
Isotype IgG  
Appearance

Liquid

Formulation

Supplied in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40% Glycerol, 0.01% Sodium azide and 0.05% BSA

Storage & Stability

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping

Shipping with blue ice.

Verification Image
ALL WB IHC-P mIHC
  • Western blot analysis was performed on extracts from 293 (lane 1, 15 μg), Hela (lane 2, 15 μg), HCT116 (lane 3, 15 μg), SW480 (lane 4, 15 μg), 3T3 (lane 5, 15 μg), and C6 (lane 6, 15 μg) using APPL Rabbit mAb.Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight.The primary antibody (1:1000 dilution) and the loading control antibody (beta-Tubulin, HY-P80955, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C.Goat Anti-Rabbit IgG-HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

  • Immunohistochemical analysis of paraffin-embedded human Ovarian Cancer‌ tissue using APPL antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82390,1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Ovarian Cancer‌ tissue using APPL antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82390,1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Liver Cancer tissue using APPL antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82390,1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Endometrial Carcinoma tissue using APPL antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82390,1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Liver Cancer tissue using APPL antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82390,1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Endometrial Carcinoma tissue using APPL antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P82390,1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Ovarian Cancer‌ tissue using APPL antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82390, 1:300 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)(HY-D1832). The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Ovarian Cancer‌ tissue using APPL antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82390, 1:300 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)(HY-D1832). The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Ovarian Cancer‌ tissue using APPL antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82390, 1:300 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)(HY-D1832). The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Liver Cancer tissue using APPL antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82390, 1:300 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)(HY-D1832). The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Liver Cancer tissue using APPL antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82390, 1:300 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)(HY-D1832). The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Liver Cancer tissue using APPL antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82390, 1:300 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)(HY-D1832). The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

Background
Function:Multifunctional adapter protein that binds to various membrane receptors, nuclear factors and signaling proteins to regulate many processes, such as cell proliferation, immune response, endosomal trafficking and cell metabolism (PubMed:10490823, PubMed:15016378, PubMed:19661063, PubMed:26073777, PubMed:26583432). Regulates signaling pathway leading to cell proliferation through interaction with RAB5A and subunits of the NuRD/MeCP1 complex (PubMed:15016378). Functions as a positive regulator of innate immune response via activation of AKT1 signaling pathway by forming a complex with APPL1 and PIK3R1 (By similarity). Inhibits Fc-gamma receptor-mediated phagocytosis through PI3K/Akt signaling in macrophages (By similarity). Regulates TLR4 signaling in activated macrophages (By similarity). Involved in trafficking of the TGFBR1 from the endosomes to the nucleus via microtubules in a TRAF6-dependent manner (PubMed:26583432). Plays a role in cell metabolism by regulating adiponecting and insulin signaling pathways (PubMed:19661063, PubMed:24879834, PubMed:26073777). Required for fibroblast migration through HGF cell signaling (By similarity). Positive regulator of beta-catenin/TCF-dependent transcription through direct interaction with RUVBL2/reptin resulting in the relief of RUVBL2-mediated repression of beta-catenin/TCF target genes by modulating the interactions within the beta-catenin-reptin-HDAC complex (PubMed:19433865)
Subcellular Localization:Early endosome membrane; Peripheral membrane protein; Nucleus; Cytoplasm; Endosome; Cell projection, ruffle; Cytoplasmic vesicle, phagosome
Expression:
Tissue_specificity:High levels in heart, ovary, pancreas and skeletal muscle
Subunit:Homodimer (PubMed:18034774). Binds RAB5A/Rab5 through an N-terminal domain. This interaction is essential for its recruitment to endosomal membranes as well as its role in cell proliferation (PubMed:15016378). Binds DCC and the catalytic domain of the inactive form of AKT2 through its PID domain (PubMed:10490823, PubMed:12011067). Binds PIK3CA and subunits of the NuRD/MeCP1 complex (PubMed:10490823, PubMed:15016378). Interacts with OCRL and INPP5B (PubMed:17765681, PubMed:20133602, PubMed:21233288, PubMed:21666675). Interacts with NTRK2 (By similarity). Interacts with APPL2; interaction is independent of follicle stimulating hormone stimulation; interaction is decreased by adiponectin in a time-dependent manner (PubMed:17030088, PubMed:18034774, PubMed:19661063). Forms a complex with APPL2 and RUVBL2 (PubMed:19433865). Forms a complex comprising APPL2, RUVBL2, CTNNB1, HDAC1 and HDAC2; interaction reduces interaction between CTNNB1, HDAC1, HDAC2 and RUVBL2 leading to the decrease of deacetylase activity of this complex; affects the recruitment of repressive complexes to the Wnt target genes (PubMed:19433865). Interacts with ANXA2 (PubMed:21645192). Interacts with TGFBR1; interaction is TGF beta dependent; mediates trafficking of the TGFBR1 from the endosomes to the nucleus via microtubules in a TRAF6-dependent manner (PubMed:26583432). Interacts with PRKCZ (PubMed:26583432). Interacts with PIK3R1 and APPL2 (By similarity). Interacts with ADIPOR1; ADIPOQ enhances this interaction; inhibites adiponectin-stimulated binding of APPL2 to ADIPOR1 (By similarity)
Database
Research Field

Cell Biology

Synonyms
APPL1; APPL; DIP13A; KIAA1428; DCC-interacting protein 13-alpha; Dip13-alpha; Adapter protein containing PH domain; PTB domain and leucine zipper motif 1
Documentation
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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APPL Antibody (YA2135)
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