Recombinant Anti-S100A8 antibody [EPR3554] (ab92331)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3554] to S100A8
- Suitable for: WB, IP, IHC-P
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-S100A8 antibody [EPR3554]
See all S100A8 primary antibodies -
Description
Rabbit monoclonal [EPR3554] to S100A8 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-Pmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human (adult and fetal) and mouse spleen tissue lysates and PMBC and HL-60 cell lysates. IHC-P: Human cervical carcinoma, breast carcinoma, tonsil and spleen tissues. IP: HL-60 cell 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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3554 -
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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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92331 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 | (4) |
1/1000. Predicted molecular weight: 11 kDa.
For unpurified use at 1/2000 - 1/10000. |
IP |
1/30.
For unpurified use at 1/20. |
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IHC-P | (1) |
1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
Notes |
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WB
1/1000. Predicted molecular weight: 11 kDa. For unpurified use at 1/2000 - 1/10000. |
IP
1/30. For unpurified use at 1/20. |
IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
Target
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Function
S100A8 is a calcium- and zinc-binding protein which plays a prominent role in the regulation of inflammatory processes and immune response. It can induce neutrophil chemotaxis and adhesion. Predominantly found as calprotectin (S100A8/A9) which has a wide plethora of intra- and extracellular functions. The intracellular functions include: facilitating leukocyte arachidonic acid trafficking and metabolism, modulation of the tubulin-dependent cytoskeleton during migration of phagocytes and activation of the neutrophilic NADPH-oxidase. Activates NADPH-oxidase by facilitating the enzyme complex assembly at the cell membrane, transferring arachidonic acid, an essential cofactor, to the enzyme complex and S100A8 contributes to the enzyme assembly by directly binding to NCF2/P67PHOX. The extracellular functions involve proinfammatory, antimicrobial, oxidant-scavenging and apoptosis-inducing activities. Its proinflammatory activity includes recruitment of leukocytes, promotion of cytokine and chemokine production, and regulation of leukocyte adhesion and migration. Acts as an alarmin or a danger associated molecular pattern (DAMP) molecule and stimulates innate immune cells via binding to pattern recognition receptors such as Toll-like receptor 4 (TLR4) and receptor for advanced glycation endproducts (AGER). Binding to TLR4 and AGER activates the MAP-kinase and NF-kappa-B signaling pathways resulting in the amplification of the proinflammatory cascade. Has antimicrobial activity towards bacteria and fungi and exerts its antimicrobial activity probably via chelation of Zn(2+) which is essential for microbial growth. Can induce cell death via autophagy and apoptosis and this occurs through the cross-talk of mitochondria and lysosomes via reactive oxygen species (ROS) and the process involves BNIP3. Can regulate neutrophil number and apoptosis by an anti-apoptotic effect; regulates cell survival via ITGAM/ITGB and TLR4 and a signaling mechanism involving MEK-ERK. Its role as an oxidant scavenger has a protective role in preventing exaggerated tissue damage by scavenging oxidants. Can act as a potent amplifier of inflammation in autoimmunity as well as in cancer development and tumor spread. The iNOS-S100A8/A9 transnitrosylase complex directs selective inflammatory stimulus-dependent S-nitrosylation of GAPDH and probably multiple targets such as ANXA5, EZR, MSN and VIM by recognizing a [IL]-x-C-x-x-[DE] motif; S100A8 seems to contribute to S-nitrosylation site selectivity. -
Tissue specificity
Calprotectin (S100A8/9) is predominantly expressed in myeloid cells. Except for inflammatory conditions, the expression is restricted to a specific stage of myeloid differentiation since both proteins are expressed in circulating neutrophils and monocytes but are absent in normal tissue macrophages and lymphocytes. Under chronic inflammatory conditions, such as psoriasis and malignant disorders, also expressed in the epidermis. Found in high concentrations at local sites of inflammation or in the serum of patients with inflammatory diseases such as rheumatoid, cystic fibrosis, inflammatory bowel disease, Crohn's disease, giant cell arteritis, cystic fibrosis, Sjogren's syndrome, systemic lupus erythematosus, and progressive systemic sclerosis. Involved in the formation and deposition of amyloids in the aging prostate known as corpora amylacea inclusions. Strongly up-regulated in many tumors, including gastric, esophageal, colon, pancreatic, bladder, ovarian, thyroid, breast and skin cancers. -
Sequence similarities
Belongs to the S-100 family.
Contains 2 EF-hand domains. -
Cellular localization
Secreted. Cytoplasm. Cytoplasm, cytoskeleton. Cell membrane. Predominantly localized in the cytoplasm. Upon elevation of the intracellular calcium level, translocated from the cytoplasm to the cytoskeleton and the cell membrane. Upon neutrophil activation or endothelial adhesion of monocytes, is secreted via a microtubule-mediated, alternative pathway. - Information by UniProt
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Database links
- Entrez Gene: 6279 Human
- SwissProt: P05109 Human
- SwissProt: P27005 Mouse
- Unigene: 416073 Human
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Alternative names
- Calgranulin-A antibody
- Calprotectin L1L subunit antibody
- CFAG antibody
see all
Images
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All lanes : Anti-S100A8 antibody [EPR3554] (ab92331) at 1/20000 dilution (purified)
Lane 1 : HL-60 (Human promyelocytic leukemia cell line) cell lysate
Lane 2 : Human fetal spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling S100A8 with purified ab92331 at 1/500.
Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).
Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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ab92331 (purified) at 1/30 immunoprecipitating S100A8 in HL-60 (Human promyelocytic leukemia cell line) cell lysate (Lane 1).
Lane 2 - rabbit monoclonal IgG instead of ab92331 in HL-60 lysates. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1500).
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling S100A8 with unpurified ab92331.
Detection: DAB staining.
Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.
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Anti-S100A8 antibody [EPR3554] (ab92331) at 1/2000 dilution (purified) + Mouse spleen tissue lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling S100A8 with unpurified ab92331. Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.
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All lanes : Anti-S100A8 antibody [EPR3554] (ab92331) (unpurified)
Lane 1 : Human spleen tissue lysate
Lane 2 : PBMC cell lysate
Lane 3 : HL-60 (Human promyelocytic leukemia cell line) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 11 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labeling S100A8 with unpurified ab92331.
Detection: DAB staining.
Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labeling S100A8 with unpurified ab92331. Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (43)
ab92331 has been referenced in 43 publications.
- Hou X et al. 3D-SeboSkin Model for Human ex vivo Studies of Hidradenitis Suppurativa/Acne Inversa. Dermatology 238:236-243 (2022). PubMed: 34077928
- Joshi A et al. Neutrophil-Derived Protein S100A8/A9 Alters the Platelet Proteome in Acute Myocardial Infarction and Is Associated With Changes in Platelet Reactivity. Arterioscler Thromb Vasc Biol 42:49-62 (2022). PubMed: 34809447
- Biswas AK et al. Targeting S100A9-ALDH1A1-Retinoic Acid Signaling to Suppress Brain Relapse in EGFR-Mutant Lung Cancer. Cancer Discov 12:1002-1021 (2022). PubMed: 35078784
- Zhang P et al. Targeting myeloid derived suppressor cells reverts immune suppression and sensitizes BRAF-mutant papillary thyroid cancer to MAPK inhibitors. Nat Commun 13:1588 (2022). PubMed: 35332119
- Liu Y et al. S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression. Cells 11:N/A (2022). PubMed: 36429008