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, Pig
    预测可用于: Sheep, Horse, Cow, Cat, 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

图片

  • 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.

    See Abreview

  • 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.

文献

This product has been referenced in:
  • Tang HL  et al. Toll-Like Receptor 4 in Paraventricular Nucleus Mediates Visceral Hypersensitivity Induced by Maternal Separation. Front Pharmacol 8:309 (2017). ICC/IF ; Mouse . Read more (PubMed: 28611665) »
  • Zou Y  et al. Effects of feeding untreated, pasteurized and acidified waste milk and bunk tank milk on the performance, serum metabolic profiles, immunity, and intestinal development in Holstein calves. J Anim Sci Biotechnol 8:53 (2017). IHC-P ; Cow . Read more (PubMed: 28630706) »

See all 59 Publications for this product

客户评价及客户问答

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Uterus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
No
Specification
Uterus
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 31 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cultured Cells (BEAS-2B Epithelial cells)
Permeabilization
Yes - 0.5% saponin
Specification
BEAS-2B Epithelial cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Selvarani Vimalanathan

Verified customer

提交于 Aug 02 2017

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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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