Name: Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y]
Brand: RabMAb
SKU: ab51253
Rating: 5 (7 reviews)

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Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP1536Y] to Alpha-synuclein (phospho S129)
Suitable for: IHC-FrFl, WB, Dot blot, ELISA, IHC-P
Reacts with: Mouse, Rat, Human

Product name

Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y]
See all Alpha-synuclein primary antibodies
Description

Rabbit monoclonal [EP1536Y] to Alpha-synuclein (phospho S129)
Host species

Rabbit
Specificity

This antibody only detects alpha synuclein phosphorylated on Ser129. IHC-P: This antibody showed no staining in human hippocampus normal brain and showed staining in Parkinson's brain as expected.

Tested Applications & Species

Application	Species
Dot	Human
IHC-FrFl	Mouse
IHC-P	Human
WB	Mouse Rat

See all applications and species data

Immunogen

Synthetic peptide within Human Alpha-synuclein aa 100 to the C-terminus. The exact sequence is proprietary.
Database link: P37840
(Peptide available as ab188826)
Positive control
- IHC-P: Human Parkinson Substantia Nigra tissue. WB: Sarkosyl-insoluble brain extract from mice transgenic for PrPA53T alpha-synuclein; Recombinant alpha-synuclein phosphorylated at S129; Rat and mouse brain lysates. IHC-Fl: Mouse brain tissue. ELISA: Alpha-synuclein (pS129) phospho peptide. Dot Blot: Human Alpha-synuclein (pS129) peptide.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP1536Y
Isotype

IgG
Research areas

Alternative Versions
- Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] - BSA and Azide free (ab209422)
Immunizing Peptide (Blocking)
- Alpha-synuclein (phospho S129) peptide (ab188826)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Alpha-synuclein protein aggregate (Active) (ab218819)
- Recombinant Human Alpha-synuclein protein (ab51189)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab51253 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
Dot	Human
IHC-FrFl	Mouse
IHC-P	Human
WB	Mouse Rat
All applications	Cow Pig Chimpanzee Macaque monkey Gorilla Orangutan Spider monkey

Application	Abreviews	Notes
IHC-FrFl	(1)	Use at an assay dependent concentration.
WB	(4)	1/1000 - 1/5000. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).Can be blocked with Alpha-synuclein (phospho S129) peptide (ab188826). Good results have been obtained by treating the membrane with 0.4% PFA for 30 min at room temperature before blocking it with 5% milk.
Dot blot		Use at an assay dependent concentration.
ELISA		Use at an assay dependent concentration.
IHC-P		Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Notes
IHC-FrFl Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).Can be blocked with Alpha-synuclein (phospho S129) peptide (ab188826). Good results have been obtained by treating the membrane with 0.4% PFA for 30 min at room temperature before blocking it with 5% milk.
Dot blot Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Application notes

Is unsuitable for Flow Cyt,IHC-Fr or IP.

Function

May be involved in the regulation of dopamine release and transport. Induces fibrillization of microtubule-associated protein tau. Reduces neuronal responsiveness to various apoptotic stimuli, leading to a decreased caspase-3 activation.
Tissue specificity

Expressed principally in brain but is also expressed in low concentrations in all tissues examined except in liver. Concentrated in presynaptic nerve terminals.
Involvement in disease

Genetic alterations of SNCA resulting in aberrant polymerization into fibrils, are associated with several neurodegenerative diseases (synucleinopathies). SNCA fibrillar aggregates represent the major non A-beta component of Alzheimer disease amyloid plaque, and a major component of Lewy body inclusions. They are also found within Lewy body (LB)-like intraneuronal inclusions, glial inclusions and axonal spheroids in neurodegeneration with brain iron accumulation type 1.
Parkinson disease 1
Parkinson disease 4
Dementia Lewy body
Sequence similarities

Belongs to the synuclein family.
Domain

The 'non A-beta component of Alzheimer disease amyloid plaque' domain (NAC domain) is involved in fibrils formation. The middle hydrophobic region forms the core of the filaments. The C-terminus may regulate aggregation and determine the diameter of the filaments.
Post-translational
modifications

Phosphorylated, predominantly on serine residues. Phosphorylation by CK1 appears to occur on residues distinct from the residue phosphorylated by other kinases. Phosphorylation of Ser-129 is selective and extensive in synucleinopathy lesions. In vitro, phosphorylation at Ser-129 promoted insoluble fibril formation. Phosphorylated on Tyr-125 by a PTK2B-dependent pathway upon osmotic stress.
Hallmark lesions of neurodegenerative synucleinopathies contain alpha-synuclein that is modified by nitration of tyrosine residues and possibly by dityrosine cross-linking to generated stable oligomers.
Ubiquitinated. The predominant conjugate is the diubiquitinated form.
Acetylation at Met-1 seems to be important for proper folding and native oligomeric structure.
Cellular localization

Cytoplasm, cytosol. Membrane. Nucleus. Cell junction, synapse. Secreted. Membrane-bound in dopaminergic neurons.
Target information above from: UniProt accession P37840 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 461375 Chimpanzee
- Entrez Gene: 101146366 Gorilla
- Entrez Gene: 6622 Human
- Entrez Gene: 20617 Mouse
- Entrez Gene: 100190854 Orangutan
- Entrez Gene: 641350 Pig
- Entrez Gene: 29219 Rat
- Omim: 163890 Human
- SwissProt: P61145 Chimpanzee
- SwissProt: P61140 Gorilla
- SwissProt: P37840 Human
- SwissProt: O55042 Mouse
- SwissProt: P61146 Orangutan
- SwissProt: P37377 Rat
- Unigene: 21374 Human
- Unigene: 17484 Mouse
- Unigene: 1827 Rat
see all
Alternative names
- Alpha synuclein antibody
- Alpha-synuclein antibody
- Alpha-synuclein, isoform NACP140 antibody
- alphaSYN antibody
- MGC105443 antibody
- MGC110988 antibody
- MGC127560 antibody
- MGC64356 antibody
- NACP antibody
- Non A beta component of AD amyloid antibody
- Non A4 component of amyloid antibody
- Non A4 component of amyloid precursor antibody
- Non-A beta component of AD amyloid antibody
- Non-A-beta component of alzheimers disease amyloid , precursor of antibody
- Non-A4 component of amyloid precursor antibody
- Non-A4 component of amyloid, precursor of antibody
- OTTHUMP00000218549 antibody
- OTTHUMP00000218551 antibody
- OTTHUMP00000218552 antibody
- OTTHUMP00000218553 antibody
- OTTHUMP00000218554 antibody
- PARK 1 antibody
- PARK 4 antibody
- PARK1 antibody
- PARK4 antibody
- Parkinson disease (autosomal dominant, Lewy body) 4 antibody
- Parkinson disease familial 1 antibody
- SNCA antibody
- Snca synuclein antibody
- Snca synuclein, alpha (non A4 component of amyloid precursor) antibody
- SYN antibody
- Synuclein alpha antibody
- Synuclein alpha 140 antibody
- Synuclein, alpha (non A4 component of amyloid precursor) antibody
- SYUA_HUMAN antibody
see all

Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

Lane 1 : Rat brain lysates, the membrane was incubated with alkaline phosphatase.
Lane 2 : Rat brain lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution

Predicted band size: 14 kDa
Observed band size: 18 kDa
why is the actual band size different from the predicted?

Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: 20 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

IHC image of alpha Synuclein (phospho S129) staining Human Parkinson Substantia Nigra tissue section*, previously antigen was retrieved by heat mediated with citrate buffer pH 6, fixed in formalin and embedded in paraffin. This section was incubated with ab51253 at 10 µg/mL for 15 mins at room temperature and detected using an HRP conjugated compact polymer system, performed on a Leica Bond™ system using the standard protocol F. DAB was used as the chromogen. Counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay performed on Human normal Substantia Nigra.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

Lane 1 : Mouse brain lysates, the membrane was incubated with alkaline phosphatase
Lane 2 : Mouse brain lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution

Predicted band size: 14 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?

Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: 40 seconds
Immunohistochemistry - Free Floating - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)This image is taken from Machado de Oliveira. R et al. PLoS Biol. 2017 Mar; 15(3): e2000374. S7a doi: 10.1371/journal.pbio.2000374 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

Expression of WT aSyn in the SN is toxic over time. Mouse brain sections immunostained for TH (red panels) and aSyn (using ab51253) (green panels) 1, 2 and 3 weeks after injection with vectors encoding for EGFP or WT aSyn. Scale bar for isolated channels 200 μm and for merged channels 100 μm.
Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)This image is courtesy of Professor Michael Goedert’s team (Sophie Morgan and Masami Masuda-Suzukake).

All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

Lane 1 : Sarkosyl-insoluble brain extract from mice transgenic for PrPA53T alpha-synuclein (line M83)
Lane 2 : Recombinant alpha-synuclein phosphorylated at S129, 3ng

Predicted band size: 14 kDa
ELISA - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

Direct ELISA antibody dose-response curve using ab51253.

Antibody concentration of 0-5000 ng/mL.

Antigen (Alpha-synuclein (pS129) phospho peptide and Alpha-synuclein non-phospho peptide) concentration of 1000 ng/mL.

An alkaline phosphatase conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Dot Blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

Dot blot analysis of human alpha Synuclein (pS129) peptide (Lane 1), huamn alpha Synuclein (unmodified) peptide (Lane 2) labelling alpha Synuclein (pS129) with ab51253 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)This image is taken from Migdalska-Richards A. et al. Ann Neurol. 2016 Nov; 80(5): 766–775. Fig 5 doi: 10.1002/ana.24790

α-Synuclein (α-syn) and S129-phosphorylated α-synuclein protein levels in SNCA/SNCA mouse brains after 12 days of treatment with 4mM ambroxol (Amb). (A) Western blotting for α-synuclein (using ab1903) and serine 129 (S129)-phosphorylated α-synuclein protein (using ab51253) in the brainstem (example blots shown).
Immunohistochemistry - Free Floating - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)This image is courtesy of an abreview by Sophie Morgan

IHC image of alpha Synuclein (phospho S129) staining in free floating tgM83^+/- or tgM83^+/+ mouse brain tissue. The section was incubated with ab51253, 1/5000, for 15 hours at 4^oC and detected using a Biotinylated conjugated Anti-Rabbit monocloal antibody, 1/200.
See Abreview
Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

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.

Click here to view the general protocols

Publishing research using ab51253? Please let us know so that we can cite the reference in this datasheet.

ab51253 has been referenced in 151 publications.

Mezias C et al. Neural connectivity predicts spreading of alpha-synuclein pathology in fibril-injected mouse models: Involvement of retrograde and anterograde axonal propagation. Neurobiol Dis 134:104623 (2020). PubMed: 31628991
Lau A et al. a-Synuclein strains target distinct brain regions and cell types. Nat Neurosci 23:21-31 (2020). PubMed: 31792467
Williams GP et al. T cell infiltration in both human multiple system atrophy and a novel mouse model of the disease. Acta Neuropathol 139:855-874 (2020). PubMed: 31993745
Wang X et al. a-synuclein promotes progression of Parkinson's disease by upregulating autophagy signaling pathway to activate NLRP3 inflammasome. Exp Ther Med 19:931-938 (2020). PubMed: 32010254
Köppen J et al. Amyloid-Beta Peptides Trigger Aggregation of Alpha-Synuclein In Vitro. Molecules 25:N/A (2020). PubMed: 32013170

View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A

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1-10 of 12 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (H4 cells)

Loading amount

5 µg

Specification

H4 cells

Gel Running Conditions

Reduced Denaturing (4-20%Tris-Glycine gel)

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 03 2009

Application

Immunohistochemistry (PFA perfusion fixed frozen sections)

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Antigen retrieval step

None

Sample

Human Tissue sections (PD patient brain sections, mesencephalon, 40 um)

Specification

PD patient brain sections, mesencephalon, 40 um

Permeabilization

Yes - 0.5% Triton-X 100

Fixative

Paraformaldehyde

Miss. Oldriska Chutna

Verified customer

Submitted May 30 2014

Application

Western blot

Loading amount

20 µg

Gel Running Conditions

Reduced Denaturing (12% TGX gel Biorad)

Sample

Human Cell lysate - whole cell (neuronal SHY5)

Specification

neuronal SHY5

Treatment

okadaic acid for 6-8 hours 37 oC

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Yien Kuo

Verified customer

Submitted Aug 30 2013

Application

Immunohistochemistry free floating

Sample

Mouse Tissue (Brain sections)

Specification

Brain sections

Sophie Morgan

Verified customer

Submitted May 23 2018

Application

Western blot

Sample

Human Tissue lysate - other (Human brain lysate)

Gel Running Conditions

Reduced Denaturing (4-12% BisTris gel)

Loading amount

1 µg

Specification

Human brain lysate

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Sophie Morgan

Verified customer

Submitted May 16 2018

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cultured Cells (HEK 293T cells)

Permeabilization

Yes - 0.1% Tritonx-100

Specification

HEK 293T cells

Blocking step

BSA as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 22°C

Fixative

Paraformaldehyde

Sophie Morgan

Verified customer

Submitted Mar 01 2018

Application

Western blot

Sample

Mouse Tissue lysate - whole (brain)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

70 µg

Treatment

alpha synuclein

Specification

brain

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 23 2015

Question

1) Could this product recognize rat samples or not though it is written "Does not react with Rat" on your data sheet?

2) What rat samples did you test?

Thank you.

Abcam community

Verified customer

Asked on Jun 25 2013

Answer

1)Our initial testing with rat showed negative and that the antibody is not recommended for use in rat. However, based on sequence analysis, it shares 100% identity with rat.

2)We previously tested rat brain, heart, kidney and spleen tissues lyates.

尚志村口

Abcam Scientific Support

Answered on Jun 25 2013

Question

We acctually do have problems with your antibody, but it's from a different batch (Lot.No. GR93844). We tried both Western blot and immunofluorecens (whole primary neurons grown on coverslips) but so far nothing has worked. A collegue of ours here at Lund University recommended this antibody and the plan is to next week try some of his batch to see if it works or if it's our samples that are the problem. Please let me know if others have problem with the same batch we got or we will continue to test it asap.

Abcam community

Verified customer

Asked on Oct 12 2012

Answer

Thank you for contacting us.

I am sorry to hear of the trouble which you have been experiencing when lot no. GR93844. We have not seen this issue with other lots of ab51253. This product is covered by our Abpromise guarantee. We are happy provide scientific support at any time. If you are using the product in species and applications listed on the datasheet and contact us within six months of purchase, we are also happy to replace or refund the product should we not be able to help you to resolve the issue.

I do hope that you are able to resolve whether this is an antibody or sample issue.If you find that the issue is with the antibody please do not hesitate to contact our Scientific Support group at technical@abcam.com who will be very happy to help you resolve any issue with this product.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

Abcam Scientific Support

Answered on Oct 12 2012

Question

Bonjour,

Je vous remercie pour votre réponse qui explicite les soucis que nous avons rencontrés récemment.

Nous avons eu ce problème de faible réactivité avec les 2 derniers tubes, ainsi qu’un autre reçu précédemment.

D’autre part, il y avait eu également une rupture de stock dû à des problèmes de production et/ou qualité l’année dernière qui avait perturbé durablement notre activité de recherche avec cet anticorps sur lequel nous comptions.

En conclusion, nous souhaiterions pouvoir bénéficier de l’envoi gratuit de 3 tubes, car il est plus compliqué de disposer d’avoir au niveau de la comptabilité.

Nous formulons le souhait d’une qualité stable des prochains lots de cet anticorps, que nous allons sans doute être obligés de vérifier à réception.

En cas de variation de sensibilité de l’anticorps observé dans votre laboratoire, nous serions intéressés d’avoir cette information en amont, avant de le découvrir par nous même.

Bien cordialement,

Abcam community

Verified customer

Asked on Jun 26 2012

Answer

Merci pour votre réponse.

Nous sommes très désolé de cette inconvenance. Veuillez croire que la qualité de nos produits est très important pour nous. J'ai transmis votre feedback aussi au laboratoire.

Nous avons actuellement trois autres tube du même lot en stock. Est-ce que ce lot vous convient et est-ce que vous souhaitez trois tubes de celui-ci? Autrement, je peux aussi vous envoyer des anticorps alternatives, contre une autre protéine,si vous souhaitez. Vous avez le droit à trois tubes d'anticorps et c'est votre choix lesquels vous souhaitez.

Veuillez ne pas hésiter à me recontacter si vous avez des questions. Je me réjouis de votre réponse

Abcam Scientific Support

Answered on Jun 26 2012

1-10 of 12 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Immunizing Peptide (Blocking)

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (151)

Customer reviews and Q&As

Western blot abreview for Anti-alpha Synuclein (phospho S129) antibody [EP1536Y]

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-alpha Synuclein (phospho S129) antibody [EP1536Y]

Western blot abreview for Anti-alpha Synuclein (phospho S129) antibody [EP1536Y]

Immunohistochemistry free floating abreview for Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y]

Western blot abreview for Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y]

Western blot abreview for Anti-alpha Synuclein (phospho S129) antibody [EP1536Y]

Post-translational
modifications