Recombinant Anti-IL-1 alpha antibody [EPR25263-27] (ab300499)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25263-27] to IL-1 alpha
- Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-IL-1 alpha antibody [EPR25263-27]
See all IL-1 alpha primary antibodies -
Description
Rabbit monoclonal [EPR25263-27] to IL-1 alpha -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details -
Species reactivity
Reacts with: Mouse
Does not react with: Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: RAW264.7 treated with 10µg/ml lipopolysaccharide, whole cell lysate. ICC/IF: RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophages), THP-1 (human monocytic leukemia monocytes). IHC-P: Mouse lung treated with lipopolysaccharides. Flow Cyt (Intra): RAW264.7 cells treated with 10ug/ml LPS, mouse bone marrow cells treated with 5ug/ml LPS.
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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: 7.20
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
EPR25263-27 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab300499 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.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/50.
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Notes |
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WB
1/1000. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/500. |
ICC/IF
1/50. |
Target
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Function
Produced by activated macrophages, IL-1 stimulates thymocyte proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast growth factor activity. IL-1 proteins are involved in the inflammatory response, being identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin and collagenase from synovial cells. -
Sequence similarities
Belongs to the IL-1 family. -
Domain
The similarity among the IL-1 precursors suggests that the amino ends of these proteins serve some as yet undefined function. -
Cellular localization
Secreted. The lack of a specific hydrophobic segment in the precursor sequence suggests that IL-1 is released by damaged cells or is secreted by a mechanism differing from that used for other secretory proteins. - Information by UniProt
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Database links
- Entrez Gene: 16175 Mouse
- SwissProt: P01582 Mouse
- Unigene: 15534 Mouse
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Alternative names
- BAF antibody
- FAF antibody
- Hematopoietin 1 antibody
see all
Images
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All lanes : Anti-IL-1 alpha antibody [EPR25263-27] (ab300499) at 1/1000 dilution
Lane 1 : Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : RAW264.7 treated with 10µg/ml lipopolysaccharide (LPS) for 7 hours, whole cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 30 kDa why is the actual band size different from the predicted?
Exposure time: 59 secondsBlocking / Diluting buffer and concentration: 5% NFDM/TBST
The expression level of IL-1 alpha was upregulated by LPS stimulation (PMID: 25870118)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1 alpha antibody [EPR25263-27] (AB300499)
Immunohistochemical analysis of paraffin-embedded Mouse lung treated tissue labeling IL-1 alpha with ab300499 at 1/500 dilution (1.05 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on mouse lung induced by LPS+BFA (A) and no staining on untreated mouse lung (B). The section was incubated with ab300499 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labeling IL-1 alpha with ab300499 at 1/50 (10.5 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 µg/mL dilution (Green). Confocal image showing increased nuclear staining in THP-1 cells treated with Phorbol-12-myristate-13-acetate (50 nM) for 24 h. is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2µg/mL dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labeling IL-1 alpha with ab300499 at 1/50 (10.5 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2µg/mL dilution (Green). Confocal image showing increased nuclear staining in Raw 264.7 cells treated with lipopolysaccharide (10 µg/ml) for 24 h (PMID:25870118) is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2µg/mL dilution.
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Flow Cytometry (Intracellular) analysis of mouse bone marrow cells treated with 5 µg/ml LPS for 7 hours (Right) / Untreated control (Left) labelling IL-1 alpha with ab300499 at a 1/500 dilution. Cells were fixed with 2% paraformaldehyde and permeabilized using True-Nuclear buffer. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, (ab150081)) was used as the secondary antibody at a 1/2000 dilution. Cells were surface stained with BV421 conjugated CD11b, then fixed with PFA followed by intracellularly stained with ab300499.
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Flow Cytometry (Intracellular) analysis of RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 10 µg/ml LPS for 24 hours (Right) / Untreated control (Left) IL-1 alpha with ab300499 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, (ab150081)) was used as the secondary antibody at a 1/2000 dilution. The expression level of IL-1 alpha was upregulated by LPS stimulation (PMID: 25870118).
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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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab300499 has not yet been referenced specifically in any publications.