Overview

  • Product name
  • Description
    Rabbit polyclonal to TLR4
  • Host species
    Rabbit
  • 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. We have obtained both positive and negative feedback from researchers using this antibody with rat samples (see Abreviews). Due to the inconsistency, we have removed rat as a guaranteed species and welcome any further feedback from researchers using this antibody.
  • Tested applications
    Suitable for: IHC-P, WB, ELISA, ICC/IF, ICC, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human TLR4 aa 420-435.
    Sequence:

    GLEQLEHLDFQ HSNLK


    Database link: O00206

  • Positive control
    • overexpressed recombinant protein

Properties

Applications

Our Abpromise guarantee covers the use of ab13556 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
IHC-P 1/50.
WB 1/500.

Use primary at 1/500 dlution in 1xTBS. Incubate at 4C for 18 hours. Secondary should be incubated at 1 hour at room temperature.

ELISA Use at an assay dependent concentration. PubMed: 24952384
ICC/IF Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • 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
    modifications
    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.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • Anti-TLR4 antibody (ab13556) at 1/500 dilution + TLR4 transfected Baculovirus-Insect whole cell lysate at 10 µg

    Secondary
    Anti-Rabbit HRP conjugate at 1/2000 dilution

    Exposure time: 6 minutes
  • ab13556 staining TLR4 in human mouse spleen tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

  • ab13556 positively staining formaldehyde fixed rat primary microglia cells (1/50), used in conjunction with goat anti rabbit Alex Fluor® 488 (1/250). 
    This image is courtesy of an Abreview submitted on 3 October 2005. We do not have any further information relating to this image.

  • ab13556 diluted 1/100 detecting TLR4 transfected mouse CHO cells by flow cytometry.   The cells were prepared by treatment with collagenase and incubated with the primary antibody for 1 hour at 22°C.  An Alexa Fluor® 488 goat anti-rabbit was used as the secondary antibody.  Cells gated on live.

    See Abreview

  • ab13556 at 1/20000 dilution staining TLR4 in mouse colitis colon tissue section by Immunohistochemistry (Bouin's fixative fixed paraffin-embedded tissue sections). Tissue underwent heat mediated antigen retrieval in microwave with two, 5 minutes incubation intervals in citrate buffer. An antibody amplifier™ was used for staining. A HRP-conjugated goat polyclonal to rabbit IgG was used as secondary at 1/10 dilution.

  • Flow cytometry analysis of Human monocytic Leukemia cells (THP-1). Cells were fixed with 2% Formaldehyde for 10 minutes at room temperature. Ab13556 was used at  2 µg/106 cells for 60 minutes at 37°C (green). Goat Anti- Rabbit Dylight 488 was used as a secondary antibody at 1:200 dilution for 40minutes at 37°C. Isotype control was Rabbit IgG under the same conditions (blue).

References

This product has been referenced in:
  • Göttle P  et al. Rescuing the negative impact of human endogenous retrovirus envelope protein on oligodendroglial differentiation and myelination. Glia 67:160-170 (2019). Read more (PubMed: 30430656) »
  • Yang WS  et al. Ectodomain Shedding of RAGE and TLR4 as a Negative Feedback Regulation in High-Mobility Group Box 1-Activated Aortic Endothelial Cells. Cell Physiol Biochem 51:1632-1644 (2018). Read more (PubMed: 30497069) »
See all 68 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-10 of 18 Abreviews

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (PFA fixed Brain)
Specification
PFA fixed Brain

Abcam user community

Verified customer

Submitted Feb 16 2019

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

Abcam user community

Verified customer

Submitted Oct 17 2017

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Cynomolgus Monkey Tissue lysate - other (macaque retina)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
300 µg
Specification
macaque retina
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 06 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cynomolgus Monkey Tissue sections (Macaque retina)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Specification
Macaque retina
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 02 2016

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Mouse Tissue sections (frozen sections from 13.5dpc mouse embryo and plac)
Specification
frozen sections from 13.5dpc mouse embryo and plac
Permeabilization
No
Fixative
Acetone

Mrs. Ira Rostovsky

Verified customer

Submitted Feb 16 2015

Application
Immunohistochemistry (Frozen sections)
Sample
Marmoset (common) Tissue sections (Caudate)
Specification
Caudate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 21 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skin)
Specification
Skin
Fixative
Acetone
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Apr 30 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (spinal cord)
Permeabilization
No
Specification
spinal cord
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative
Formaldehyde

Ms. Kelly Atkins

Verified customer

Submitted Oct 08 2010

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (spinal cord)
Specification
spinal cord
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jul 30 2010

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Rat spinal cord homogenate)
Gel Running Conditions
Reduced Denaturing (7.5% SDS-PAGE)
Loading amount
40 µg
Treatment
Sham or L5 nerve transection
Specification
Rat spinal cord homogenate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted May 30 2008

1-10 of 18 Abreviews

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

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