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    tlr4-antibody-ab13556.pdf

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Immunology Innate Immunity Macrophage / Inflamm.
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Anti-TLR4 antibody (ab13556)

  • Datasheet
  • SDS
Reviews (19)Q&A (26)References (151)

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Western blot - Anti-TLR4 antibody (ab13556)
  • Western blot - Anti-TLR4 antibody (ab13556)
  • Immunohistochemistry (Frozen sections) - Anti-TLR4 antibody (ab13556)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR4 antibody (ab13556)
  • Flow Cytometry - Anti-TLR4 antibody (ab13556)
  • Immunohistochemistry (Frozen sections) - Anti-TLR4 antibody (ab13556)
  • Flow Cytometry - Anti-TLR4 antibody (ab13556)

Key features and details

  • Rabbit polyclonal to TLR4
  • Suitable for: WB, IHC-P, IHC-Fr, Flow Cyt
  • Reacts with: Mouse, Human, Recombinant fragment
  • Isotype: IgG

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Overview

  • Product name

    Anti-TLR4 antibody
    See all TLR4 primary antibodies
  • 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 & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Mouse
    WB
    Mouse
    Recombinant fragment
    See all applications and species data
  • Immunogen

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

    GLEQLEHLDFQ HSNLK


    Database link: O00206
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 99% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Immunology
    • Immunoglobulins
    • Receptors
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Immunology
    • Innate Immunity
    • TLR Signaling

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human TLR4 protein (Active) (ab233665)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-P
Mouse
WB
Mouse
Recombinant fragment
Application Abreviews Notes
WB (5)
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.

IHC-P (2)
Use at an assay dependent concentration.
IHC-Fr (5)
Use at an assay dependent concentration.
Flow Cyt (3)
Use at an assay dependent concentration.

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

Notes
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.

IHC-P
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.
  • Target information above from: UniProt accession O00206 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7099 Human
    • Entrez Gene: 21898 Mouse
    • Omim: 603030 Human
    • SwissProt: O00206 Human
    • SwissProt: Q9QUK6 Mouse
    • Unigene: 174312 Human
    • Unigene: 38049 Mouse
    • 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

    • Western blot - Anti-TLR4 antibody (ab13556)
      Western blot - Anti-TLR4 antibody (ab13556)
      Anti-TLR4 antibody (ab13556) at 1/1000 dilution + partial recombinant mouse TLR4 protein, 100 ng
    • Western blot - Anti-TLR4 antibody (ab13556)
      Western blot - Anti-TLR4 antibody (ab13556)
      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
    • Immunohistochemistry (Frozen sections) - Anti-TLR4 antibody (ab13556)
      Immunohistochemistry (Frozen sections) - Anti-TLR4 antibody (ab13556)

      ab13556 at a 1/100 dilution staining TLR4 in mouse spleen tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR4 antibody (ab13556)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR4 antibody (ab13556)

      ab13556 at a 1/100 dilution staining TLR4 in mouse spleen tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    • Flow Cytometry - Anti-TLR4 antibody (ab13556)
      Flow Cytometry - Anti-TLR4 antibody (ab13556)This image is courtesy of an Abreview submitted by Dr alexandre garin

      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

    • Immunohistochemistry (Frozen sections) - Anti-TLR4 antibody (ab13556)
      Immunohistochemistry (Frozen sections) - Anti-TLR4 antibody (ab13556)

      ab13556 at 1/100000 dilution (12 hrs at 4degC) staining TLR4 in mouse colitis colon tissue section by Immunohistochemistry (Formalin-fixed tissue sections). A biotin Goat Anti-Rabbit secondary was used at 1/2000 for 1 hour at RT. Counterstain: Methyl Green at 200µL for 2 mins at RT. Localization: Inflammatory cells.

    • Flow Cytometry - Anti-TLR4 antibody (ab13556)
      Flow Cytometry - Anti-TLR4 antibody (ab13556)

      Flow cytometry analysis of THP-1 (Human monocytic leukemia cell line). 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 40 minutes at 37°C. Isotype control was Rabbit IgG under the same conditions (blue).

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (151)

    Publishing research using ab13556? Please let us know so that we can cite the reference in this datasheet.

    ab13556 has been referenced in 151 publications.

    • Shi CX  et al. Sevoflurane attenuates brain damage through inhibiting autophagy and apoptosis in cerebral ischemia-reperfusion rats. Mol Med Rep 21:123-130 (2020). PubMed: 31746402
    • Kang X  et al. The TLR4/ERK/PD-L1 axis may contribute to NSCLC initiation. Int J Oncol 57:456-465 (2020). PubMed: 32468028
    • Jing N  et al. Exogenous activation of cannabinoid-2 receptor modulates TLR4/MMP9 expression in a spinal cord ischemia reperfusion rat model. J Neuroinflammation 17:101 (2020). PubMed: 32248810
    • Tong W  et al. Resveratrol inhibits LPS-induced inflammation through suppressing the signaling cascades of TLR4-NF-?B/MAPKs/IRF3. Exp Ther Med 19:1824-1834 (2020). PubMed: 32104238
    • Yang H  et al. Procyanidin Compound (PC) Suppresses Lipopolysaccharide-Induced Cervical Cancer Cell Proliferation Through Blocking the TLR4/NF-?B Pathway. Cancer Manag Res 12:497-509 (2020). PubMed: 32158256
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 45 Abreviews or Q&A

    Western blot abreview for Anti-TLR4 antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (tumor)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    30 µg
    Specification
    tumor
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 10 2020

    Immunohistochemistry free floating abreview for Anti-TLR4 antibody

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Mouse Tissue sections (PFA fixed Brain)
    Specification
    PFA fixed Brain
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 16 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-TLR4 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 17 2017

    Western blot abreview for Anti-TLR4 antibody

    Below Average
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 06 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-TLR4 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 02 2016

    Immunohistochemistry (Frozen sections) abreview for Anti-TLR4 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mrs. Ira Rostovsky

    Verified customer

    Submitted Feb 16 2015

    Immunohistochemistry (Frozen sections) abreview for Anti-TLR4 antibody

    Inconclusive
    Abreviews
    Abreviews
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 21 2014

    Question

    what is being used as antibody amplifier(TM)
    How was the image obtained?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 01 2012

    Answer

    Thank you for contacting us.

    I have received a response from the lab:

    The antibody amplifier(TM) refers to a platform that was used. I have requestedthis productto be added to our catalog. I will let you know if this is possible.

    The product description says:

    The Antibody Amplifier™ is an apparatus for IHC and IF that utilizes a submersion method to saturate mounted tissue on slides with extremely diluted antibody. The Antibody Amplifier™ rocks slides on a standard laboratory rocker during antibody incubation steps. It contains 12 watertight wells, each capable of holding a single standard microscope slide in 3 mls of reagent. It is covered with a snap-on lid. It overcomes the typical problems that occur in IHC and IF, mainly uneven staining, lack of reproducibility, and high antibody costs. Antibody Amplifier™ boxes can be stacked on a rocker; they should be taped together for further stability.

    Thus, the antibody amplifier is a way to incubate antibodies, not an amplification step (ABC method or polymer) as I had thought.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Aug 01 2012

    Question

    Is there a blocking peptide available for this antibody?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 23 2012

    Answer

    We do not sell a blocking peptide for this product unfortunately.

    Sorry I couldn't be more helpful here. Please contact us again if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Jul 23 2012

    Question

    Thanks for your reply. I would like to receive a replacement product. Please send me the TLR4 antibody (cat#22048). The purchase order number for the previous one is xxxx My shipping address: xxxxx

    Read More

    Abcam community

    Verified customer

    Asked on Jul 23 2012

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement to the address you have provided with the order number xxxxx. This should arrive tomorrow morning.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Abcam Scientific Support

    Answered on Jul 23 2012

    1-10 of 45 Abreviews or Q&A

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