Anti-TLR4 antibody [76B357.1] (ab22048)
Key features and details
- Mouse monoclonal [76B357.1] to TLR4
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, ICC, IHC-Fr, ELISA
- Reacts with: Mouse, Rat, Human, Pig
- Isotype: IgG2b
Overview
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Product name
Anti-TLR4 antibody [76B357.1]
See all TLR4 primary antibodies -
Description
Mouse monoclonal [76B357.1] to TLR4 -
Host species
Mouse -
Specificity
TLR4 expression levels and cleavage or degradation products can vary between different cell and tissue samples. Customers have observed this variability in WB band size and our laboratory has confirmed this variability as well observing lower molecular weight cleavage and degradation products and in some samples a lack of the full length TLR4 band. The TLR4 cleavage and degradation products and potential lack of full length TLR4 are well documented in the literature, including PMID 16885150 and 22927440. We recommend running a positive control human intestine tissue lysate. -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, ICC, IHC-Fr, ELISAmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Pig
Predicted to work with: Sheep, Horse, Cow, Cat, Chimpanzee -
Immunogen
Synthetic peptide corresponding to Human TLR4 aa 100-200 conjugated to Keyhole Limpet Haemocyanin (KLH).
Database link: O00206 -
Positive control
- Human, Mouse and Rat small intestine for Western Blot or THP1 cells for FACS analysis. Flow Cyt (Intra): Jurkat cells.
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General notes
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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: 7.40
Preservative: 0.05% Sodium azide
Constituents: 99% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
76B357.1 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Buffer
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab22048 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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Flow Cyt (Intra) |
Use 1µg for 106 cells.
Methanol or paraformaldehyde fixed cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (5) |
Use at an assay dependent concentration.
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IHC-P | (5) |
Use at an assay dependent concentration.
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ICC | (3) |
Use at an assay dependent concentration.
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IHC-Fr | (2) |
Use at an assay dependent concentration.
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ELISA |
Use at an assay dependent concentration. PubMed: 24952384
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Notes |
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Flow Cyt (Intra)
Use 1µg for 106 cells. Methanol or paraformaldehyde fixed cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. |
ICC
Use at an assay dependent concentration. |
IHC-Fr
Use at an assay dependent concentration. |
ELISA
Use at an assay dependent concentration. PubMed: 24952384 |
Target
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Function
Cooperates with LY96 and CD14 to mediate the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MYD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Also involved in LPS-independent inflammatory responses triggered by Ni(2+). These responses require non-conserved histidines and are, therefore, species-specific. -
Tissue specificity
Highly expressed in placenta, spleen and peripheral blood leukocytes. Detected in monocytes, macrophages, dendritic cells and several types of T-cells. -
Involvement in disease
Genetic variation in TLR4 is associated with age-related macular degeneration type 10 (ARMD10) [MIM:611488]. ARMD is a multifactorial eye disease and the most common cause of irreversible vision loss in the developed world. In most patients, the disease is manifest as ophthalmoscopically visible yellowish accumulations of protein and lipid that lie beneath the retinal pigment epithelium and within an elastin-containing structure known as Bruch membrane. -
Sequence similarities
Belongs to the Toll-like receptor family.
Contains 18 LRR (leucine-rich) repeats.
Contains 1 LRRCT domain.
Contains 1 TIR domain. -
Domain
The TIR domain mediates interaction with NOX4. -
Post-translational
modificationsN-glycosylated. Glycosylation of Asn-526 and Asn-575 seems to be necessary for the expression of TLR4 on the cell surface and the LPS-response. Likewise, mutants lacking two or more of the other N-glycosylation sites were deficient in interaction with LPS. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 281536 Cow
- Entrez Gene: 7099 Human
- Entrez Gene: 21898 Mouse
- Entrez Gene: 399541 Pig
- Entrez Gene: 29260 Rat
- Omim: 603030 Human
- SwissProt: Q9GL65 Cow
- SwissProt: Q9MYW3 Horse
see all -
Alternative names
- ARMD10 antibody
- CD284 antibody
- CD284 antigen antibody
see all
Images
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Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling TLR4 with ab22048 at 5 ug/ml. The section was blocked using blocking solution (1% BSA in PBS) for 1 hour at room temperature and incubated with ab22048 overnight at 4°C after removing blocking solution.
Antigen retrieval: Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).
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ab22048 staining TLR4 in Human stomach tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 1 hour; antigen retrieval was by heat mediation in citrate buffer (10mM, pH 6) (ab64236). Samples were incubated with primary antibody (1/100) for 1 hour at 23°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.
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Overlay histogram showing Jurkat cells (ab7899) stained with ab22048 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum (ab7481) / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22048, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.
Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback. -
Representative examples of immunohistochemical staining of TLR4 in colorectal carcinoma tissues (original magnification 100×).
Paraffin-embedded human normal colorectal tissue (A) and colorectal carcinoma tissue (D) stained for TLR4 with ab22048 in immunohistochemical analysis. Counter stained with hematoxylin.
No staining is observed in normal colorectal tissue (A).
(From Figure 4A and 4D of Xu et al)
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ab22048 (5 ug/ml) staining TLR4 in Human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells and the adjacent keratinocytes in the epidermal layer.
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ab22048 at 5 µg/ml staining TLR4 in Human colon tissue sections by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). Left hand image shows staining with an isotype control antibody whilst right one shows staining with ab22048.
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ab22048 staining TLR4 in Rat's salivary gland tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). ab22048 at a dilution of 1:100 generated a membrane-cytoplasmic staining in the tissue with stronger signal in ductal epithelial cells.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (261)
ab22048 has been referenced in 261 publications.
- Zheng Z et al. E. coli JM83 damages the mucosal barrier in Ednrb knockout mice to promote the development of Hirschsprung‑associated enterocolitis via activation of TLR4/p‑p38/NF‑κB signaling. Mol Med Rep 25:N/A (2022). PubMed: 35302172
- Li S et al. The Protective Effects of Ginseng Polysaccharides and Their Effective Subfraction against Dextran Sodium Sulfate-Induced Colitis. Foods 11:N/A (2022). PubMed: 35327312
- Liu L et al. The transmembrane adapter SCIMP recruits tyrosine kinase Syk to phosphorylate Toll-like receptors to mediate selective inflammatory outputs. J Biol Chem 298:101857 (2022). PubMed: 35337798
- Zheng H et al. Cordyceps militaris Modulates Intestinal Barrier Function and Gut Microbiota in a Pig Model. Front Microbiol 13:810230 (2022). PubMed: 35369439
- Qu P et al. Application Value of Serum Hcy, TLR4, and CRP in the Diagnosis of Cerebral Small Vessel Disease. Evid Based Complement Alternat Med 2022:4025965 (2022). PubMed: 35502170