Anti-TLR4抗体[76B357.1] (ab22048)

概述

  • 产品名称Anti-TLR4抗体[76B357.1]
    参阅全部 TLR4 一抗
  • 描述
    小鼠单克隆抗体[76B357.1] to TLR4
  • 特异性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.
  • 经测试应用适用于: ELISA, IHC-P, IHC-Fr, Flow Cyt, ICC, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Sheep, Horse, Cow, Cat, Pig, Chimpanzee
  • 免疫原

    Synthetic peptide corresponding to Human TLR4 aa 100-200 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: O00206

  • 阳性对照
    • Human, Mouse and Rat small intestine for Western Blot or THP1 cells for FACS analysis. Jurkat cells.

性能

应用

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.

应用 Ab评论 说明
ELISA Use at an assay dependent concentration. PubMed: 24952384
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Antigen retrieval with citrate buffer pH 6.
IHC-Fr 1/100.
Flow Cyt 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 Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

靶标

  • 功能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.
  • 组织特异性Highly expressed in placenta, spleen and peripheral blood leukocytes. Detected in monocytes, macrophages, dendritic cells and several types of T-cells.
  • 疾病相关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.
  • 序列相似性Belongs to the Toll-like receptor family.
    Contains 18 LRR (leucine-rich) repeats.
    Contains 1 LRRCT domain.
    Contains 1 TIR domain.
  • 结构域The TIR domain mediates interaction with NOX4.
  • 翻译后修饰N-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.
  • 细胞定位Membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ARMD10 antibody
    • CD284 antibody
    • CD284 antigen antibody
    • Homolog of Drosophila toll antibody
    • hToll antibody
    • TLR 4 antibody
    • TLR4 antibody
    • TLR4_HUMAN antibody
    • TOLL antibody
    • Toll like receptor 4 antibody
    • Toll-like receptor 4 antibody
    see all

Anti-TLR4 antibody [76B357.1] 图像

  • 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.

    See Abreview

  • ab22048 staining TLR4 in Mouse spleen tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% serum for 30 minutes at 20°C. Samples were incubated with primary antibody (1/100 in PBS-BSA) for 1 hour at 20°C. An Alexa Fluor®488-conjugated Rat anti mouse IgG polyclonal (1/200) was used as the secondary antibody.

    TLR4 labelling (yellow) co-localises with CD35 labelling (red) on follicular dendritic cells in the spleen.  TLR 4 alone (green) can also be seen on other cell types in the spleen- most probably macrophages.

    See Abreview

  • 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.

  • Immunofluorescence analysis of murine macrophages, staining TLR4 with ab22048.

    Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 10 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 1% BSA) for 12 hours at 4°C. A TexasRed®-conjugated donkey anti-mouse (ab6818) polyclonal IgG (1/100) was used as the secondary antibody.

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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.

  • 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.

Anti-TLR4 antibody [76B357.1] (ab22048)参考文献

This product has been referenced in:
  • Li G  et al. TLR4-mediated NF-?B signaling pathway mediates HMGB1-induced pancreatic injury in mice with severe acute pancreatitis. Int J Mol Med 37:99-107 (2016). WB ; Mouse . Read more (PubMed: 26719855) »
  • Guo J  et al. LPS/TLR4-mediated stromal cells acquire an invasive phenotype and are implicated in the pathogenesis of adenomyosis. Sci Rep 6:21416 (2016). IHC-P, ICC/IF ; Human . Read more (PubMed: 26898650) »

See all 30 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - whole (kidney)
Gel Running Conditions Non-reduced Denaturing (8%)
Loading amount 10 µg
Specification kidney
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Feb 02 2016

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (liver)
Specification liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Dec 01 2014

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Sample Mouse Tissue sections (follicular dendritic cells in spleen)
Specification follicular dendritic cells in spleen
Permeabilization No
Fixative Acetone
Username

Ms. Laura McCulloch

Verified customer

提交于 Oct 15 2013

The Flow Cytometry image using ab22048 was obtained with secondary antibody product ab47041: Goat polyclonal Secondary Antibody to Mouse IgG - H&L (PE), pre-adsorbed.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Macrophages)
Specification Macrophages
Fixative Formaldehyde
Permeabilization Yes - .1% TritonX-100 in 3% BSA
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Dr. Varkha Agrawal

Verified customer

提交于 Oct 18 2012

Application Western blot
Sample Mouse Cell lysate - whole cell (NIH3T3 fibroblast, N2a neuronal cell, primary macr)
Loading amount 0.2 µg
Specification NIH3T3 fibroblast, N2a neuronal cell, primary macr
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Mikael Rutberg

Verified customer

提交于 Sep 05 2012

Suitable positive controls include human, mouse or rat small intestine samples for WB applications, and THP1 or Jurkat cells for Flow Cytometry applications.

For use in WB applications, small intestine whole cell lysate products from either human (ab29276) or mouse (ab79390 are suitable positive controls.
For use in IHC applications, we can recommend product ab4327: Colon (Human) Normal Tissue Slides (a...

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Overall, human and sheep TLR4 proteins share only 75% homology. For the sheep TLR4 reference sequence, please see UniProt #B5DC89: http://www.uniprot.org/uniprot/B5DC89.

However, the synthetic peptide immunogen used to generate ab22048 shows ...

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In this case, the higher MW bands (~170kDa) very likely represent the heavily glycosylated or complexed form or TLR4. The lower MW band (~95kDa) represents TLR4.

To break down any complexes further, it may be beneficial to heat the samples f...

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