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:
  • Wu XJ  et al. Penehyclidine hydrochloride inhibits TLR4 signaling and inflammation, and attenuates blunt chest trauma and hemorrhagic shock-induced acute lung injury in rats. Mol Med Rep 17:6327-6336 (2018). Read more (PubMed: 29488614) »
  • Cao W  et al. Homeobox a5 Promotes White Adipose Tissue Browning Through Inhibition of the Tenascin C/Toll-Like Receptor 4/Nuclear Factor Kappa B Inflammatory Signaling in Mice. Front Immunol 9:647 (2018). Read more (PubMed: 29651293) »
See all 59 Publications for this product

Customer reviews and Q&As

1-10 of 43 Abreviews or Q&A

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

Question
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

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Question
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.

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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.

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Answer

Thank you for contacting Abcam.

I am very sorry to hear about the troubles which you have been experiencing using this product and wish to thank you for providing the details of your protocol.

Our products are covered by Abpromise guarantee for a period of six months after delivery. Should this product not perform as indicated on the datasheet, we provide scientific support, replacement or refund. More information on our Abpromise may be found at the following link: https://www.abcam.com/index.html?pageconfig=abpromise

I would be happy to provide you with refund or replacement if this product has been purchased within the last six months. In order to provide this could you please send me the Abcam order confirmation number or PO used to purchase ab13556? Would it also be preferable to you for me to send ab22048 as a replacement or a new lot of ab13556?

I look forward to your reply. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Answer

Thank you very much for your interest in our TLR4 antibodies.
Based on the homology between the Sheep sequence (UniProt: B5DC89) and their immunogens, I would recommend ab47093 or ab13556, both of which are 94% homologous. Our antibody ab22048 also shows 93% homology to Sheep TLR4.
Because none of these antibodies has been validated for use in sheep, I can offer a discount off a future purchase if you buy one of the antibodies now, test it in sheep and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.
If you are interested in this offer, please follow these steps:
1. Reply to this e-mail to let me know which antibody you would like to proceed and test in sheep. I will then send a discount code. This code must be issued before purchasing the antibody so please wait for my reply before ordering.
2. Purchase the antibody either by phone, fax, or online (www.abcam.com).
3. Test it in sheep.
4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.
5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.
We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody turns out to be unsuitable for sheep, you will still receive the discount on your next purchase after your Abreview has been submitted.
Please let me know if you have any questions about this offer and I would be happy to help you further.
The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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1-10 of 43 Abreviews or Q&A

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

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