Anti-TLR4 antibody [76B357.1] ab22048 is a mouse monoclonal antibody that is used in TLR4 IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.
- Antibody clone 76B357.1 has been tried and trusted by researchers since 2006 and is cited in >490 publications
IgG2b
Mouse
pH: 7.4
Preservative: 0.05% Sodium azide
Constituents: 99% PBS, 0.05% BSA
Liquid
Monoclonal
ICC | ELISA | IHC-P | ICC/IF | IHC-Fr | Flow Cyt (Intra) | |
---|---|---|---|---|---|---|
Human | Expected | Expected | Tested | Expected | Expected | Tested |
Mouse | Expected | Expected | Predicted | Expected | Expected | Predicted |
Rat | Expected | Expected | Tested | Expected | Expected | Expected |
Cat | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Chimpanzee | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Cow | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Horse | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Pig | Expected | Expected | Predicted | Expected | Expected | Predicted |
Sheep | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Pig, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Horse, Cow, Cat, Chimpanzee | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Pig, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Horse, Cow, Cat, Chimpanzee | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Horse, Cow, Cat, Chimpanzee, Mouse, Pig | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Pig, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Horse, Cow, Cat, Chimpanzee | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Pig, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Horse, Cow, Cat, Chimpanzee | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/cells for 6.00000 µg/cells | Notes Methanol or paraformaldehyde fixed cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Horse, Cow, Cat, Chimpanzee, Mouse, Pig | Dilution info - | Notes - |
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Cooperates with LY96 and CD14 to mediate the innate immune response to bacterial lipopolysaccharide (LPS) (PubMed:27022195). Acts via MYD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response (PubMed:9237759, PubMed:10835634, PubMed:27022195,PubMed:21393102). Also involved in LPS-independent inflammatory responses triggered by free fatty acids, such as palmitate, and Ni(2+). Responses triggered by Ni(2+) require non-conserved histidines and are, therefore, species-specific (PubMed:20711192). Both M.tuberculosis HSP70 (dnaK) and HSP65 (groEL-2) act via this protein to stimulate NF-kappa-B expression (PubMed:15809303). In complex with TLR6, promotes sterile inflammation in monocytes/macrophages in response to oxidized low-density lipoprotein (oxLDL) or amyloid-beta 42. In this context, the initial signal is provided by oxLDL- or amyloid-beta 42-binding to CD36. This event induces the formation of a heterodimer of TLR4 and TLR6, which is rapidly internalized and triggers inflammatory response, leading to the NF-kappa-B-dependent production of CXCL1, CXCL2 and CCL9 cytokines, via MYD88 signaling pathway, and CCL5 cytokine, via TICAM1 signaling pathway, as well as IL1B secretion. Binds electronegative LDL (LDL(-)) and mediates the cytokine release induced by LDL(-) (PubMed:23880187). Stimulation of monocytes in vitro with M.tuberculosis PstS1 induces p38 MAPK and ERK1/2 activation primarily via TLR2, but also partially via this receptor (PubMed:16622205, PubMed:10835634, PubMed:15809303, PubMed:17478729, PubMed:20037584, PubMed:20711192, PubMed:23880187, PubMed:27022195, PubMed:9237759). Activated by the signaling pathway regulator NMI which acts as damage-associated molecular patterns (DAMPs) in response to cell injury or pathogen invasion, therefore promoting nuclear factor NF-kappa-B activation (PubMed:29038465).
Toll-like receptor 4, hToll, TLR4
Anti-TLR4 antibody [76B357.1] ab22048 is a mouse monoclonal antibody that is used in TLR4 IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.
- Antibody clone 76B357.1 has been tried and trusted by researchers since 2006 and is cited in >490 publications
IgG2b
Mouse
pH: 7.4
Preservative: 0.05% Sodium azide
Constituents: 99% PBS, 0.05% BSA
Liquid
Monoclonal
76B357.1
Affinity purification Protein G
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.
kappa
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
Toll-like receptor 4 (TLR4) is a membrane protein involved in the innate immune response. It serves as a receptor that recognizes pathogen-associated molecular patterns such as lipopolysaccharides from bacterial cell walls. Often referred to by its alternate names TLR-2 and CD284 this protein has a molecular weight of approximately 95 kDa. TLR4 is mainly expressed on the surface of immune cells like macrophages and dendritic cells enabling these cells to detect and respond to microbial invaders effectively.
TLR4 plays an important role in activating the immune system. It forms a complex with myeloid differentiation factor 2 (MD-2) to facilitate binding to its ligands. Upon activation TLR4 triggers a signaling cascade that leads to the release of pro-inflammatory cytokines and interferons which are important for initiating the immune response. This early detection mechanism alerts the immune system to the presence of pathogens allowing the body to mount a defense.
TLR4 participates in the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. These pathways are central to the inflammatory response with TLR4 acting as a sensor that activates downstream components like the MyD88 and TRIF proteins. These interactions result in the expression of genes that produce cytokines and enzymes involved in inflammation and immunity.
TLR4 has been implicated in the development of sepsis and atherosclerosis. In sepsis TLR4 recognizes lipopolysaccharides from bacterial infections which can cause excessive inflammation if uncontrolled. In atherosclerosis TLR4 receptors on vascular cells respond to endogenous ligands such as oxidized low-density lipoproteins contributing to plaque formation. The TLR4 interaction with proteins like CD14 and MD-2 is significant as it influences the inflammatory pathways that exacerbate these conditions.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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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) (100x Citrate Buffer pH 6.0 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.
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)
Overlay histogram showing Jurkat cells (Jurkat whole cell lysate ab7899) stained with ab22048 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum (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) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (Mouse IgG2b [PLPV219] - Isotype Control 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.
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.
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.
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.
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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