Recombinant Anti-APPL antibody [EPR13569] - BSA and Azide free (ab250150)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13569] to APPL - BSA and Azide free
- Suitable for: IHC-P, ICC/IF, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-APPL antibody [EPR13569] - BSA and Azide free
See all APPL primary antibodies -
Description
Rabbit monoclonal [EPR13569] to APPL - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, MCF7 and HEK-293 cell lysates. IHC-P: Human pancreas and colon carcinoma tissue. ICC/IF: MCF7 cells.
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General notes
ab250150 is the carrier-free version of ab180140.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Affinity purified -
Clonality
Monoclonal -
Clone number
EPR13569 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab250150 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 90 kDa (predicted molecular weight: 80 kDa).
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 90 kDa (predicted molecular weight: 80 kDa). |
Target
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Function
Required for the regulation of cell proliferation in response to extracellular signals from an early endosomal compartment. Links Rab5 to nuclear signal transduction. -
Tissue specificity
High levels in heart, ovary, pancreas and skeletal muscle. -
Sequence similarities
Contains 1 PH domain.
Contains 1 PID domain. -
Domain
Overexpression of an N-terminal domain (residues 1-319) or a C-terminal region (residues 273-709) has a proapoptotic effect. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Early endosome membrane. Nucleus. Early endosomal membrane-bound and nuclear. Translocated into the nucleus upon release from endosomal membranes following internalization of EGF. - Information by UniProt
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Database links
- Entrez Gene: 26060 Human
- Entrez Gene: 72993 Mouse
- Entrez Gene: 290537 Rat
- Omim: 604299 Human
- SwissProt: Q9UKG1 Human
- SwissProt: Q8K3H0 Mouse
- Unigene: 476415 Human
- Unigene: 202322 Mouse
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Alternative names
- Adapter protein containing PH domain antibody
- Adaptor Protein Containing pH Domain antibody
- Adaptor protein containing pH domain PTB domain and leucine zipper motif 1 antibody
see all
Images
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All lanes : Anti-APPL antibody [EPR13569] (ab180140) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : APPL CRISPR/Cas9 edited HeLa cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 80 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180140).
Lanes 1- 4: Merged signal (red and green). Green - ab180140 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab180140 was shown to react with Anti-APPL in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab265187 (CRISPR/Cas9 edited cell lysate ab257836) lane below 90kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and APPL1 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180140 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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This data was developed using ab180140, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human pancreas carcinoma tissue labeling APPL with ab180140 at 1/100 dilution. Secondary antibody Goat anti rabbit IgG(HRP) prediluted. Counter stain Hematoxylin. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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All lanes : Anti-APPL antibody [EPR13569] (ab180140) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : 293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 80 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?This data was developed using ab180140, the same antibody clone in a different buffer formulation.
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This data was developed using ab180140, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling APPL with ab180140 at 1/100 dilution. Secondary antibody Goat anti rabbit IgG(HRP) prediluted. Counter stain Hematoxylin. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab250150 has not yet been referenced specifically in any publications.