Recombinant Anti-AACT antibody [EPR17088-68] (ab205198)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17088-68] to AACT
- Suitable for: WB, IHC-P, ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-AACT antibody [EPR17088-68]
See all AACT primary antibodies -
Description
Rabbit monoclonal [EPR17088-68] to AACT -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human plasma (untreated and PNGase F treated) and blood; Human fetal spleen lysate. IHC-P: Human tonsil tissue. ICC/IF: Raji cells. IP: Human plasma 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.
Properties
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17088-68 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab205198 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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WB |
1/1000. Detects a band of approximately 50-70 kDa (predicted molecular weight: 47 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/50.
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IP |
1/40.
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Notes |
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WB
1/1000. Detects a band of approximately 50-70 kDa (predicted molecular weight: 47 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/50. |
IP
1/40. |
Target
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Function
Although its physiological function is unclear, it can inhibit neutrophil cathepsin G and mast cell chymase, both of which can convert angiotensin-1 to the active angiotensin-2. -
Tissue specificity
Plasma. Synthesized in the liver. Like the related alpha-1-antitrypsin, its concentration increases in the acute phase of inflammation or infection. Found in the amyloid plaques from the hippocampus of Alzheimer disease brains. -
Involvement in disease
Defects in SERPINA3 may be a cause of chronic obstructive pulmonary disease (COPD) [MIM:107280]. -
Sequence similarities
Belongs to the serpin family. -
Domain
The reactive center loop (RCL) extends out from the body of the protein and directs binding to the target protease. The protease cleaves the serpin at the reactive site within the RCL, establishing a covalent linkage between the carboxyl group of the serpin reactive site and the serine hydroxyl of the protease. The resulting inactive serpin-protease complex is highly stable. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 12 Human
- Omim: 107280 Human
- SwissProt: P01011 Human
- Unigene: 534293 Human
- Unigene: 710488 Human
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Alternative names
- SERPINA3 antibody
- AACT antibody
- AACT_HUMAN antibody
see all
Images
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Anti-AACT antibody [EPR17088-68] (ab205198) at 1/2000 dilution + Human plasma at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID: 9880565).
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling AACT with ab205198 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab205198) at 1/500 dilution. Cytoplasmic staining on human tonsil tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (Human Burkitt's lymphoma cell line) cells labeling AACT with ab205198 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weak nuclear staining on Raji cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab205198 at 1/50 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + His-Tagged Human SERPINA3 (aa 24 to 324) recombinant protein 100ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 47 kDa
Observed band size: 33 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Diluting buffer and concentration 5% NFDM/TBST
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Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + Human blood at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 47 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + Human fetal spleen lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-AACT antibody [EPR17088-68] (ab205198) at 1/2000 dilution
Lane 1 : Untreated Human plasma
Lane 2 : Human plasma treated with PNGase F
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution
Predicted band size: 47 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?
Exposure time: 4 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
De-glycosylation treatment with PNGase, demonstrates a molecular weight shift from 50-70kDa to 46kDa. This result is consistent with what has been described in literature the literature (PMID:) 9880565.
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AACT was immunoprecipitated from 1 mg of Human plasma lysate with ab205198 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab205198 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: Human plasma lysate, 10 μg (Input).
Lane 2: ab205198 IP in Human plasma lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab205198 in Human plasma lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab205198 has been referenced in 2 publications.
- Huang K et al. Construction and validation of a glioblastoma prognostic model based on immune-related genes. Front Neurol 13:902402 (2022). PubMed: 35968275
- Fan Z et al. Immune-related SERPINA3 as a biomarker involved in diabetic nephropathy renal tubular injury. Front Immunol 13:979995 (2022). PubMed: 36304455