Anti-TLR3 抗体 [40C1285] (ab13915)

Reviews (3)Q&A (9)References (16)

製品の概要

  • 製品名

    Anti-TLR3 antibody [40C1285]
    TLR3 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [40C1285] to TLR3

  • 由来種

    Mouse

  • 特異性

    Negative results in WB have been obtained for mouse interstine lysates (human intestine is a positive control). The protein is heavily glycosylated and the observed band size should be higher than the predicted 103kDa. The protein is found at different molecular weight in different tissues so optimization may be necessary.

  • アプリケーション

    適用あり: IHC-P, WB, Flow Cyt, IPmore details

  • 種交差性

    交差種: Mouse, Human

  • 免疫原

    Synthetic peptide:

    VLNLTHNQLRRLPAAN

    conjugated to KLH, corresponding to amino acids 55-70 of Human TLR3

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • Human intestine tissue lysate, mouse spleen

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab13915 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-P Use a concentration of 5 - 10 µg/ml. Permeabilization of samples might be necessary and overnight incubation with the ab13915 is recommended.
WB Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 108 kDa). TLR3 is a single-pass type I membrane protein. We recomend not to boil the samples, instead heat lysates at 60-70C for 10 minutes.
Flow Cyt Use 2-5µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
IP Use 2µg for 106 cells.

ターゲット情報

  • 機能

    Key component of innate and adaptive immunity. TLRs (Toll-like receptors) control host immune response against pathogens through recognition of molecular patterns specific of microorganisms. TLR3 is a nucleotide-sensing TLR which is activated by double-stranded RNA, a sign of viral infection. Acts via MYD88 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response.

  • 組織特異性

    Expressed at high level in placenta and pancreas. Also detected in CD11c+ immature dendritic cells. Only expressed in dendritic cells and not in other leukocytes, including monocyte precursors. TLR3 is the TLR that is expressed most strongly in the brain, especially in astrocytes, glia, and neurons.

  • 関連疾患

    Defects in TLR3 are associated with herpes simplex encephalitis type 2 (HSE2) [MIM:613002]. HSE is a rare complication of human herpesvirus 1 (HHV-1) infection, occurring in only a small minority of HHV-1 infected individuals. HSE is characterized by hemorrhagic necrosis of parts of the temporal and frontal lobes. Onset is over several days and involves fever, headache, seizures, stupor, and often coma, frequently with a fatal outcome. Note=TLR3 mutations predispose otherwise healthy individuals to isolated herpes simplex encephalitis through a mechanism that involves impaired IFNs production and reduced immune defense against viral infection in the central nervous system.

  • 配列類似性

    Belongs to the Toll-like receptor family.
    Contains 22 LRR (leucine-rich) repeats.
    Contains 1 LRRCT domain.
    Contains 1 LRRNT domain.
    Contains 1 TIR domain.

  • ドメイン

    ds-RNA binding is mediated by LRR 1 to 3, and LRR 17 to 18.

  • 翻訳後修飾

    Heavily N-glycosylated, except on that part of the surface of the ectodomain that is involved in ligand binding.

  • 細胞内局在

    Endoplasmic reticulum membrane. Endosome membrane.
  • Information by UniProt

  • 参照データベース

    • 別名

      • CD283 antibody
      • CD283 antigen antibody
      • IIAE2 antibody
      • TLR 3 antibody
      • Tlr3 antibody
      • TLR3_HUMAN antibody
      • Toll Like Receptor 3 antibody
      • Toll-like receptor 3 antibody
      see all

    画像

    • Western blot - Anti-TLR3 antibody [40C1285] (ab13915)
      Western blot - Anti-TLR3 antibody [40C1285] (ab13915)
      All lanes : Anti-TLR3 antibody [40C1285] (ab13915) at 3 µg/ml

      Lane 1 : Untransfected HEK293 cell lysate
      Lane 2 : HEK293 cell lysate transfected with human TLR3 cDNA whole cell lysate
      Lane 3 : Human intestine tissue lysate
      Lane 4 : Human placenta tissue lysate
      Lane 5 : Human heart tissue lysate
      Lane 6 : Human ovary tissue lysate

      Secondary
      All lanes : Goat anti-mouse HRP conjugate

      Predicted band size: 108 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR3 antibody [40C1285] (ab13915)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR3 antibody [40C1285] (ab13915)

      Immunohistochemical analysis of formalin fixed paraffin embedded tissue section of mouse intestine using ab13915 at 1/500 dilution with HRP-DAB detection and hematoxylin counterstaining. The image shows a punctate staining of the ER and endosomes in a subset of cells in Peyer's patches (organized lymphoid nodules) in the tested section.

    • Flow Cytometry - Anti-TLR3 antibody [40C1285] (ab13915)
      Flow Cytometry - Anti-TLR3 antibody [40C1285] (ab13915)

      Flow cytometry analysis of human B cells using ab13915 at 1µg/106 cells followed by FITC conjugated secondary antibody (Red). Green shows isotype control.  

    • Western blot - Anti-TLR3 antibody [40C1285] (ab13915)
      Western blot - Anti-TLR3 antibody [40C1285] (ab13915)

      Western blot analysis of TLR3 in lysates from untransfected 293 cells (lane A), 293 cells transfected with human TLR3 cDNA (lane B), and 20 µg/lane human intestine tissue lysate (lane C).

      Western blot analysis of TLR3 in lysates from untransfected 293 cells (lane A), 293 cells transfected with human TLR3 cDNA (lane B), and 20 µg/lane human intestine tissue lysate (lane C).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR3 antibody [40C1285] (ab13915)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR3 antibody [40C1285] (ab13915)

      Immunohistochemical analysis of formalin fixed paraffin embedded tissue section of mouse colon using ab13915 at 1/500 dilution with HRP-DAB detection and hematoxylin counterstaining. Intense signal was found in subset of cells at the bases of the crypts.

    • Flow Cytometry - Anti-TLR3 antibody [40C1285] (ab13915)
      Flow Cytometry - Anti-TLR3 antibody [40C1285] (ab13915)

      ab13915 at 2µg / 1x106 cells staining intracellular TLR3 in human monocytes by Flow Cytometery. A PE-conjugated goat polyclonal to mouse IgG1 was used as secondary. The shaded histogram repersents monocytes without antibody; green repersents isotype control and red histogram shows staining with ab13915. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR3 antibody [40C1285] (ab13915)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR3 antibody [40C1285] (ab13915)

      ab13915 at 5ug/ml staining TLR3 in mouse spleen tissue section by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded sections). The left image repersents staining with isotype control and right one repersents staining with ab13915.

    • Flow Cytometry - Anti-TLR3 antibody [40C1285] (ab13915)
      Flow Cytometry - Anti-TLR3 antibody [40C1285] (ab13915)This image is a courtesy of Tanja Matijevic

      ab13915 staining TLR3 in SW480, SW620 (colon cancer) and FaDu, Detroit 562 obtained from human cells by Flow Cytometry. Samples were prepared by trypsinizining and washing in PBS and centrifugation in a buffer containing 2% FBS, 0.1% Na-azide and PBS. Cells were fixed in paraformaldehyde and permeabilized in 0.2% Tween. Gating and analysis was performed by WinMDI 2.9 software. The primary antibody was diluted to 1 µg/cells (PBS, 2% FBS and 0.1% Na-azide) and incubated with sample for 30 minutes at 4°C. An FITC conjugated rabbit polyclonal to mouse IgG, diluted to 1/40, was used as secondary. Image represents FACS analysis of TLR3 protein expression in SW480 (A), SW620 (B), FaDu (C) and Detroit 562 (D) cell lines. Cell lines were permeabilized and immunostained with TLR3-specific antibody (thick black l

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    参考文献

    This product has been referenced in:

    • Gong M  et al. Pattern recognition receptor-mediated innate immune responses in seminal vesicle epithelial cell and their impacts on cellular function. Biol Reprod N/A:N/A (2019). Read more (PubMed: 31350848) »
    • Zhang Z  et al. Spinal circRNA-9119 Suppresses Nociception by Mediating the miR-26a-TLR3 Axis in a Bone Cancer Pain Mouse Model. J Mol Neurosci N/A:N/A (2019). Read more (PubMed: 31368062) »
    See all 16 Publications for this product

    ab13915 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    レビューと Q&A

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    1-10 of 12 Abreviews or Q&A

    Western blot abreview for Anti-TLR3 antibody [40C1285]

     Average
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 01 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-TLR3 antibody [40C1285]

     Inconclusive
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
    Sample
    Mouse Cell (Macrophages)
    Specification
    Macrophages
    Permeabilization
    Yes - Triton X 0.1% 5 min
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Jul 23 2014

    Question

    I want to reproduce the staining I obtained in paraffin embedded sections using Ab 13915 ( anti human TLR3). I think of using Ab 12085. Can you please tell me if Ab 12085 binds to a different or same epitope as Ab13915? Or do Ab13915 and Ab12085 have the same or different paratope? Do you have other alternatives for a primary antibody? Thank you in advance! Regards,

    Read More
    Answer

    Thank you for contacting us.

    Unfortunately the epitope for these two antibodies has not been mapped, and therefore I am unable to provide the sequence they recognise.

    However, ab13915 is currently in stock. If this antibody gave you good results I would recommend you to continue to use it. Ab13915 is a very good antibody.

    We do have some other anti TLR3 antibodies validated for IHC-P. You can have a look at the list by https://www.abcam.com/Search?Keywords=TLR3+&fReset=1&pt=1&source=TopSearch&btnSearch=Search#Search?AppliedFacets.FacetProductType=Primary+antibodies&AppliedFacets.FacetTargetName=TLR3&AppliedFacets.FacetApplication=IHC-P&IsFacetsOnly=False&PageNumber=1&Keyw.

    I hope this information is helpful to you. If not please let me know and I will be happy to assist you further.

    Read More

    Question


    Inquiry: We have had the TLR3（ab13915），Please tell us if this antibody may be applied in immuohistochemistry and immunofluorescence for frozen section. Thank you very much!

    Read More
    Answer

    Thank you for contacting us. This antibody has not been tested in frozen sections or in cells, or in paraffin embedded tissue sections. Typically though, antibodies that work well in paraffin embedded sections also work in frozen tissues.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    Kindly advise in regards to the following information. The images are attached.
    Antibody code: ab13915, ab22048, ab134368
    details:
    Ab13915: Po: 89238 Lot: GR52440 Invoice: 1345585
    Ab22048: Po: 89238 Lot: GR77209 Invoice: 1345585
    Ab134368: Po: 89238 Lot: GR95073 Invoice: 1346631
    Antibody storage conditions (temperature/reconstitution etc): for ab13915 and ab22048 at -20- -80ºC ,
    for ab134368 at 4 ºC
    Description of the problem (high background, wrong band size, more bands, no band etc.): too many nonspecific band that could not allow me to determine my band
    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
    Purified proteins from human cell lines
    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
    RIPA buffer with protease inhibitors and heating the samples for 10mins at 70 ºC
    Amount of protein loaded
    Out of the transfected cell 5yg and 20ug were beingloaded, out of the cell lines 100ug were loaded
    Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
    Reducing gels. 4-12% and 3-8%
    Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
    Transfer buffer: glycin, trisma base and methanol, transfer time: 45min for one membrane. Blocking: 5% skim milk, also for ab13915 I tried3% BSA. I tried 1.5hrs blocking and I tried overnight only for ab13915
    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): Abcam, they all react with human, mouse. The antibody was diluted in PBS with 0.02% NA-azide (also I tried 3% BSA for ab13915). Dilution used was: 2ug/ml for ab13915, Dilution used was: 3ug/ml for ab22048, Dilution used was: 3ug/ml for ab134368. Incubation time was overnight at 4 ºC and also for ab13915 I tried 1.5hrs.
    Three washings for 5mins with PBST
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step):
    Dako, HRP-antimouse, diluted 1:60 in %5 blocking skim milk for 20mins, followed by three washes with PBST 5mins each with shaking
    Detection method (ECL, ECLPlus etc.): ECL
    Positive and negative controls used (please specify): for ab13915 transfected and untrasnfected cells with TLR3 plamis (pEF6/V6-His-TOPO, for ab22048 THP1 cell lines which are known to express TLR4, ab134368 transfected and untrasfected cells withTLR9 plasmid PcDNA3
    Optimization attempts (problem solving)
    How many times have you tried the Western? Three to four times
    Have you run a "No Primary" control? No
    Do you obtain the same results every time? e.g. are the background bands always in the same place? Yes
    What steps have you altered? I tried to t do overnight blocking, to do Blocking with BSA, to incubate the primary antibody for 1.5hrs only rater than overbight,..
    Thanks in advance for your assistance.
    Kind Regards,

    Read More
    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.
    I have read the details you have kindly provided and have following further suggestions;
    - Try loading 30-40 ug of protein lysates.
    - Heat the lysates for 5-10 minutes at 100C.
    - Filter the buffers before use.
    - Block the membrane with 3-5% BSA in all cases for 2 hours at room temperature.
    - Please make sure the lysates are prepared in ice cold RIPA buffer. Keep the lysates on ice.
    - Use TBST as dilution and washing buffer.
    - Try loading control with same lysates and no primary control (no primary antibody)>
    - Please check the cells should be 80-90% confluent.
    - use fresh protease inhibitors.
    I hope these suggestions would help to improve results. I will look forward to hearing from you soon.

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    Question

    This email has 2 sections:

    A recap of the complaint

    The respondent’s survey results (section-by-section)
    Recap of the complaint

    Date(s) submitted: July 9, 2012

    Catalog number(s): 13915

    Product(s): Mouse monoclonal [40C1285] to TLR3

    CCEID: 3934045
    Survey results
    Telephone section
    Q. Did you contact Abcam's Scientific Support by telephone in reference to this complaint?
    A. No
    Q. Was it clear from the phone options which number you needed to press to get your issue resolved?
    A.
    Protocol advice section
    Q. Did Abcam's Scientific Support give you protocol advice?
    A.Yes
    Q. How did you feel about being given protocol advice?
    A.I requested protocol advice
    Q. Did the product work successfully after following the advice?
    A.No
    Q. Why have you not tried the protocol advice? (Select all that apply.)
    Free of charge replacement section
    Q. Did Abcam's Scientific Support give you a free of charge replacement?
    A. No

    Q. Did the replacement product work successfully?
    Credit note / refund section
    Q. Did Abcam's Scientific Support give you a credit note or a refund?
    A.No

    Q. Would you have preferred a different outcome from receiving a credit note or refund?
    A.
    Overall satisfaction section
    Q. How satisfied are you with how the complaint was handled?
    A.Satisfied
    Q. Could we have done anything to better resolve the problem?
    A.dont know!
    Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
    A.No, I would not mind
    Q. Please provide a phone number or email address where we can reach you.
    A.WE found ourself a solution for the problem and got satisfying staining with the product.

    Read More
    Answer

    Thank you for completing the survey.

    I am glad to hear that the product worked fine at the end. Could you share the details of step you have altered for better results; it may help other scientists in community?

    Many thanks for your cooperation during the whole case.

    Best of luck with you research.

    Read More

    Question

    Product code: 13915
    Lot number: GR52440-3
    Inquiry: Despite seveal trials to stain slides from paraffinembedded colonic biopsies, we do not succeed to get any staining of TLR3 at all. We have 4 micrometer thin slides. -Heated at 60degr C for 30 min. -Deparrafination with Neo-Clear, 100%,96%, 80%,70% then rehydration and H2O2 to block endogen peroxidase. -Antigen retrieval buffer pH 6 or 9. Boiled at 15 min, then resting in room temp for 17 min. -Buffer/detergent using TBS+0.05% tween or PBS+0.25% triton 100-X. -Primary antibody diluted 1:50-1:100- 1:200, incubated 1 hour at room temp. or 24 hours at 4 degr. C. -Secondary antibody Dako Envision.(rabbit/mouse)30min room temp. -Then DAB+ for at leaset 5 min. Always wash 1, 3 and 3 times 5 min in buffer/detergent after antigen retrieval buffer, primary and sec. antibody. -Lastly hematoxylin staining. We have used this protocol and the same slides/ biopsies for several other antibodies at IHC staining with success, but this Ab is not working. Do you have any suggestions? Possible to have it refunded? Or preferrably get a new ab that works! I do not upload any slides because they are totally negative no staining at all except one slide pH 9, 1hour 1:50 diluted primary antibody with a little background staining. Kindly

    Read More
    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    The protocol seems fine to me however because antibodies needs optimization when they used with paraffin embedded sections so I have following suggestions, that might help to improve results;

    - Recommend trying antigen retrieval with microwave (700 watt full power) 5-10 minutes incubation in citrate buffer pH 6.0. or EDTA buffer pH 9.0. Please carefully check the buffer pH, in many cases the incorrect pH fails the antibodies.
    - Pressure cooker; the slides can be cooked for 2 minutes at full required pressure.
    - Could you provide the order number?

    I am sure optimizing the antigen retrieval step would help in getting good staining. However, if the results do not improve please contact me for further assistance.

    Read More

    Question

    We bought the primary antibody ab 13915; mouse monoclonal antihuman anti TLR3 antibody from you recently. Our purpose was to stain slides from colonic biopsies with histological inflammation,embedded in parrafin. We have followed a standard prosedure with deparafinization, rehydration, boiling in citrat-buffer pH6 and trisbuffer pH 9, also tried both PBS with 0.25% triton-X or TBS+ 0,05% tween. We tried incubation in 1 hour and also over night. We have been using concentration of anti TLR3 of 2.5-5 and 10microg/mL. Only when using antigen retrival buffer trisbuffer pH9 kons 10microgr/mL over night incubation we got some backgroud staining.

    We have stained slidesthe same biopsies with other primary antibodies, using the same reagens / protocoll and it does work, but thisAb 13915does not work at all!



    Do you have any idea why we do not get any staining at all ?

    Do you have any idea what to do to make it work?

    If not wehope to get the money payed back!

    Read More
    Answer

    Thank you for contacting us.

    I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


    I look forward to receiving your reply.

    Read More

    Question

    Hello,
    We have purchased 3 TLR antibodies from your company and I'd like to know if you have blocking peptides for them;
    TLR3- ab13915
    TLR7- ab45371
    TLR9- ab12121
    I'd appreciate any help!
    Thank you,

    Read More
    Answer

    Thank you for contacting us.

    We do notsell blocking peptide for ab12121 so I am sorry we will not be able to provide this.

    I have been waiting an answer from lab regarding ab13915 and ab45371; I will send you the info soon.

    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

    Question

    I have used ab62566 to detect TLR3 from a variety of tumor cell lines, including HEK293 and HepG2. These cell lines should be negative for TLR3. My positive control has a TLR3 expression plasmid transfected to produce full length protein. However I see a band at the expected molecular weight in every lane, transfected or not. the datasheet for ab13915 shows a blot illustrating what i expect to see even using 293 cells so i think this antibody is not recognizing TLR3.

    Read More
    Answer

    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 for one vial of ab13915.

    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

    1-10 of 12 Abreviews or Q&A

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