Recombinant Anti-BIN1 antibody [EPR13463] (ab182562)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13463] to BIN1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-BIN1 antibody [EPR13463]
See all BIN1 primary antibodies -
Description
Rabbit monoclonal [EPR13463] to BIN1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: A431, HeLa and U-87 MG whole cell lysate. Human brain and muscle lysate. IHC-P: Human ovarian carcinoma tissue. IP: ab182562 in HeLa whole cell lysate. Flow Cyt: HeLa cells. ICC/IF: HeLa cells.
-
General notes
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR13463 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab182562 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 |
---|---|---|
Flow Cyt (Intra) |
1/30 - 1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB |
1/1000 - 1/10000. Detects a band of approximately 65, 70 kDa (predicted molecular weight: 65 kDa).
|
|
IHC-P |
1/100 - 1/500.
Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). |
|
ICC/IF | (1) |
1/250.
|
IP |
1/20 - 1/40.
|
Notes |
---|
Flow Cyt (Intra)
1/30 - 1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Detects a band of approximately 65, 70 kDa (predicted molecular weight: 65 kDa). |
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). |
ICC/IF
1/250. |
IP
1/20 - 1/40. |
Target
-
Function
May be involved in regulation of synaptic vesicle endocytosis. May act as a tumor suppressor and inhibits malignant cell transformation. -
Tissue specificity
Ubiquitous. Highest expression in the brain and muscle. Isoform IIA is expressed only in the brain where it is concentrated in axon initial segments and nodes of Ranvier. Isoform BIN1 is widely expressed with highest expression in skeletal muscle. -
Involvement in disease
Defects in BIN1 are the cause of centronuclear myopathy autosomal recessive (ARCNM) [MIM:255200]; also known as autosomal recessive myotubular myopathy. Centronuclear myopathies are congenital muscle disorders characterized by progressive muscular weakness and wasting involving mainly limb girdle, trunk, and neck muscles. It may also affect distal muscles. Weakness may be present during childhood or adolescence or may not become evident until the third decade of life. Ptosis is a frequent clinical feature. The most prominent histopathologic features include high frequency of centrally located nuclei in muscle fibers not secondary to regeneration, radial arrangement of sarcoplasmic strands around the central nuclei, and predominance and hypotrophy of type 1 fibers. -
Sequence similarities
Contains 1 BAR domain.
Contains 1 SH3 domain. -
Post-translational
modificationsPhosphorylated by protein kinase C. -
Cellular localization
Cytoplasm and Nucleus. - Information by UniProt
-
Database links
- Entrez Gene: 274 Human
- Omim: 601248 Human
- SwissProt: O00499 Human
- Unigene: 193163 Human
-
Alternative names
- AMPH 2 antibody
- AMPH2 antibody
- Amphiphysin 2 antibody
see all
Images
-
All lanes : Anti-BIN1 antibody [EPR13463] (ab182562) at 1/5000 dilution
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 65 kDaExposure times: Lane 1: 180 seconds; Lane 2: 20 seconds; Lane 3: 10 seconds.
Blocking/Dilution buffer: 5%NFDM/TBST.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue sections labeling BIN1 with purified ab182562 at 1/500 dilution (0.52 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
-
All lanes : Anti-BIN1 antibody [EPR13463] (ab182562) at 1/10000 dilution (Purified)
Lane 1 : Human brain lysates
Lane 2 : Human muscle lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 65 kDa
Observed band size: 50-65 kDa why is the actual band size different from the predicted?The multiple bands are isoforms produced by alternative splicing
-
ab182562 (purified) at 1/20 dilution (1 µg) immunoprecipitating BIN1 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab182562 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab182562 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BIN1 with purified ab182562 at 1/30 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
-
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BIN1 with purified ab182562 at 1:250 dilution (1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (8)
ab182562 has been referenced in 8 publications.
- Lambert E et al. The Alzheimer susceptibility gene BIN1 induces isoform-dependent neurotoxicity through early endosome defects. Acta Neuropathol Commun 10:4 (2022). PubMed: 34998435
- Fagg WS et al. Definition of germ layer cell lineage alternative splicing programs reveals a critical role for Quaking in specifying cardiac cell fate. Nucleic Acids Res 50:5313-5334 (2022). PubMed: 35544276
- Marques-Coelho D et al. Differential transcript usage unravels gene expression alterations in Alzheimer's disease human brains. NPJ Aging Mech Dis 7:2 (2021). PubMed: 33398016
- Young AMH et al. A map of transcriptional heterogeneity and regulatory variation in human microglia. Nat Genet 53:861-868 (2021). PubMed: 34083789
- Karlina R et al. Identification and characterization of distinct brown adipocyte subtypes in C57BL/6J mice. Life Sci Alliance 4:N/A (2021). PubMed: 33257475
- Chen C et al. RNA-seq analysis of the key long noncoding RNAs and mRNAs related to cognitive impairment after cardiac arrest and cardiopulmonary resuscitation. Aging (Albany NY) 12:14490-14505 (2020). PubMed: 32693388
- De Rossi P et al. Neuronal BIN1 Regulates Presynaptic Neurotransmitter Release and Memory Consolidation. Cell Rep 30:3520-3535.e7 (2020). PubMed: 32160554
- Ge Y et al. The splicing factor RBM25 controls MYC activity in acute myeloid leukemia. Nat Commun 10:172 (2019). PubMed: 30635567