Recombinant Anti-AP2S1 antibody [EPR2697(2)] (ab128950)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2697(2)] to AP2S1
- Suitable for: WB, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-AP2S1 antibody [EPR2697(2)]
See all AP2S1 primary antibodies -
Description
Rabbit monoclonal [EPR2697(2)] to AP2S1 -
Host species
Rabbit -
Specificity
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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Tested Applications & Species
Application Species IHC-P HumanIP HumanWB MouseHuman -
Immunogen
Synthetic peptide within Human AP2S1 aa 100-200. The exact sequence is proprietary.
Database link: P53680 -
Positive control
- WB: Human fetal brain, SH-SY5Y, HEK-293T, HeLa, Mouse brain, Rat brain lysates, Mouse kidney and Mouse brain lysates. IHC-P: Human breast carcinoma, thyroid carcinoma and colon carcinoma tissues IP: Human fetal brain lysate
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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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2697(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab128950 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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IHC-P |
Human
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IP |
Human
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WB |
Mouse
Human
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Application | Abreviews | Notes |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 17 kDa.
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IP |
1/10 - 1/100.
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 17 kDa. |
IP
1/10 - 1/100. |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
Target
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Function
Component of the adaptor protein complex 2 (AP-2). Adaptor protein complexes function in protein Transport via Transport vesicles in different membrane traffic pathways. Adaptor protein complexes are vesicle coat components and appear to be involved in cargo selection and vesicle formation. AP-2 is involved in clathrin-dependent endocytosis in which cargo proteins are incorporated into vesicles surrrounded by clathrin (clathrin-coated vesicles, CCVs) which are destined for fusion with the early endosome. The clathrin lattice serves as a mechanical scaffold but is itself unable to bind directly to membrane components. Clathrin-associated adaptor protein (AP) complexes which can bind directly to both the clathrin lattice and to the lipid and protein components of membranes are considered to be the major clathrin adaptors contributing the CCV formation. AP-2 also serves as a cargo receptor to selectively sort the membrane proteins involved in receptor-mediated endocytosis. AP-2 seems to play a role in the recycling of synaptic vesicle membranes from the presynaptic surface. AP-2 recognizes Y-X-X-[FILMV] (Y-X-X-Phi) and [ED]-X-X-X-L-[LI] endocytosis signal motifs within the cytosolic tails of transmembrane cargo molecules. AP-2 may also play a role in maintaining normal post-endocytic trafficking through the ARF6-regulated, non-clathrin pathway. The AP-2 alpha and AP-2 sigma subunits are thought to contribute to the recognition of the [ED]-X-X-X-L-[LI] motif. -
Sequence similarities
Belongs to the adaptor complexes small subunit family. -
Cellular localization
Cell membrane. Membrane > coated pit. AP-2 appears to be excluded from internalizing CCVs and to disengage from sites of endocytosis seconds before internalization of the nascent CCV. - Information by UniProt
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Database links
- Entrez Gene: 1175 Human
- Entrez Gene: 232910 Mouse
- Entrez Gene: 65046 Rat
- Omim: 602242 Human
- SwissProt: P53680 Human
- SwissProt: P62743 Mouse
- SwissProt: P62744 Rat
- Unigene: 119591 Human
see all -
Alternative names
- Adapter-related protein complex 2 sigma subunit antibody
- Adaptor protein complex AP 2 subunit sigma antibody
- Adaptor protein complex AP-2 subunit sigma antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2S1 antibody [EPR2697(2)] (ab128950)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human thyroid carcinoma tissue sections labeling AP2S1 with purified ab128950 at 1:100 dilution (12.07 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-AP2S1 antibody [EPR2697(2)] (ab128950) at 1/1000 dilution (Purified)
Lane 1 : Human fetal brain lysates
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 4 : Mouse brain lysates
Lane 5 : Mouse kidney lysates
Lane 6 : Rat brain 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: 17 kDa
Observed band size: 17 kDa14% gel is preferred to 4-20% gradient gel
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Anti-AP2S1 antibody [EPR2697(2)] (ab128950) at 1/1000 dilution (Purified) + Mouse kidney lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 17 kDa
Observed band size: 17 kDa -
All lanes : Anti-AP2S1 antibody [EPR2697(2)] (ab128950) at 1/5000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : Mouse brain lysates
Lane 3 : Rat brain lysates
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: 17 kDa
Observed band size: 17 kDa -
ab128950 (purified) at 1:60 dilution (2µg) immunoprecipitating AP2S1 in Human fetal brain lysate.
Lane 1 (input): Human fetal brain lysate 10µg
Lane 2 (+): ab128950 & Human fetal brain lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab128950 in Human fetal brain lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
All lanes : Anti-AP2S1 antibody [EPR2697(2)] (ab128950) at 1/1000 dilution
Lane 1 : Fetal brain lysate
Lane 2 : SH-SY5Y lysate
Lane 3 : 293T lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 17 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2S1 antibody [EPR2697(2)] (ab128950)ab128950, at a 1/250 dilution, staining AP2S1 in paraffin embedded Human breast carcinoma tissue by Immunohistochemistry.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2S1 antibody [EPR2697(2)] (ab128950)ab128950, at a 1/250 dilution, staining AP2S1 in paraffin embedded Human colon carcinoma tissue by Immunohistochemistry.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2S1 antibody [EPR2697(2)] (ab128950)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon carcinoma tissue sections labeling AP2S1 with purified ab128950 at 1:100 dilution (12.07 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab128950 has been referenced in 5 publications.
- Pascolutti R et al. Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling. Cell Rep 27:3049-3061.e6 (2019). PubMed: 31167147
- Beacham GM et al. NECAPs are negative regulators of the AP2 clathrin adaptor complex. Elife 7:N/A (2018). PubMed: 29345618
- Kadlecova Z et al. Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2. J Cell Biol 216:167-179 (2017). PubMed: 28003333
- Aguet F et al. Membrane dynamics of dividing cells imaged by lattice light-sheet microscopy. Mol Biol Cell 27:3418-3435 (2016). PubMed: 27535432
- Biase FH et al. Cell fate inclination within 2-cell and 4-cell mouse embryos revealed by single-cell RNA sequencing. Genome Res 24:1787-96 (2014). ICC ; Mouse . PubMed: 25096407