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Anti-Bone Sialoprotein抗体(ab52128)

Reviews (1)Q&A (5)References (36)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bone Sialoprotein antibody (ab52128)

    Key features and details

    • Rabbit polyclonal to Bone Sialoprotein
    • Suitable for: IHC-P
    • Reacts with: Human
    • Isotype: IgG

    概述

    • 产品名称

      Anti-Bone Sialoprotein抗体
      参阅全部 Bone Sialoprotein 一抗

    • 描述

      兔多克隆抗体to Bone Sialoprotein

    • 宿主

      Rabbit

    • 经测试应用

      适用于: IHC-Pmore details

    • 种属反应性

      与反应: Human

    • 免疫原

      Human Bone Sialoprotein (bone extract)

    • 常规说明

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    性能

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab52128于以下的经测试应用

    “应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

    应用 Ab评论 说明
    IHC-P
    Use at an assay dependent concentration.
    说明
    IHC-P
    Use at an assay dependent concentration.

    靶标

    • 功能

      Binds tightly to hydroxyapatite. Appears to form an integral part of the mineralized matrix. Probably important to cell-matrix interaction. Promotes Arg-Gly-Asp-dependent cell attachment.

    • 翻译后修饰

      N-glycosylated; glycans consist of sialated and core-fucosylated bi-, tri- and tetraantennary chains.
      O-glycosylated at eight sites; mucin-type glycans contain Gal, GlcNAc, GalNAc and terminal NeuAc.
      Sulfated on either Tyr-313 or Tyr-314.

    • 细胞定位

      Secreted.
    • Information by UniProt

    • 数据库链接

      • 别名

        • BNSP antibody
        • Bone sialoprotein 2 antibody
        • Bone sialoprotein II antibody
        • BSP antibody
        • BSP II antibody
        • BSPII antibody
        • Cell binding sialoprotein antibody
        • Cell-binding sialoprotein antibody
        • IBSP antibody
        • Integrin binding sialoprotein antibody
        • Integrin-binding sialoprotein antibody
        • SIAL_HUMAN antibody
        • SPII antibody
        see all

      图片

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bone Sialoprotein antibody (ab52128)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bone Sialoprotein antibody (ab52128)
        ab52128 (2ug/ml) staining Bone Sialoprotein in human bone using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of Bone sialoprotein in sectretory compartments in the cells of the perichondrium
        Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

      文献 (36)

      发表研究结果有使用 ab52128？请让我们知道，以便我们可以引用本数据表中的参考文章。

      ab52128 被引用在 36 文献中.

      • Cheng C  et al. Repair of a Rat Mandibular Bone Defect by Hypertrophic Cartilage Grafts Engineered From Human Fractionated Adipose Tissue. Front Bioeng Biotechnol 10:841690 (2022). PubMed: 35350180
      • Huang RL  et al. Engineering hypertrophic cartilage grafts from lipoaspirate for critical-sized calvarial bone defect reconstruction: An adipose tissue-based developmental engineering approach. Bioeng Transl Med 7:e10312 (2022). PubMed: 36176620
      • Zhou T  et al. Conditioned medium derived from 3D tooth germs: A novel cocktail for stem cell priming and early in vivo pulp regeneration. Cell Prolif 54:e13129 (2021). PubMed: 34585454
      • Engel N  et al. An optimized 3D-printed perfusion bioreactor for homogeneous cell seeding in bone substitute scaffolds for future chairside applications. Sci Rep 11:22228 (2021). PubMed: 34782672
      • Wang N  et al. miR-6807-5p Inhibited the Odontogenic Differentiation of Human Dental Pulp Stem Cells Through Directly Targeting METTL7A. Front Cell Dev Biol 9:759192 (2021). PubMed: 34790668
      View all Publications for this product

      客户评价及客户问答

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      1-6 of 6 Abreviews or Q&A

      Immunohistochemistry (Frozen sections) abreview for Anti-Bone Sialoprotein antibody

       Below Average
      Abreviews
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (Endochondral bone)
      Specification
      Endochondral bone
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - 0.1% Triton
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 19°C
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      提交于 Jan 25 2011

      Question


      I'm sorry for the delay in answering you, but the costumer was in doubt about what option to take. Now she decided.

      She will accept the ab125227 free of charge as replacement.

      Thanks for all your help.

      Best Regards,

      Read More
      Answer

      Thank you very much for your reply.

      The replacement vial of ab125227 will ship out with our next delivery to Biogen. Due to Customs restrictions, this replacement will ship out at the full price ($303.30) but I have arranged credit note # **** to reimburse you for the price.

      Please keep me updated about how this replacement antibody works for the customer, and let me know if you have any questions or need anything else from us.

      Read More

      Question

      We did the test accordingly to your instructions:

      Antigen Retrieval: Pre-heated Citrate Buffer pH 6,0 (95-99oC). Water-Bath for 30 minutes.

      Peroxidase Blocking: Hydrogen Peroxide Block Reagent (Spring – Code DHP-125), for 20 minutes, at room temperature (20-25oC).

      Protein Blocking: Protein Block Reagent (Spring - Code DBP-125), for 10 minutes, at room temperature.

      Primary Antibody: Diluted 1:500 in Antibody Diluent (Spring - Code ADS-125). Overnight Incubation (16 hours), at 4oC.

      Detection System: Polymer-HRP N-Histofine Simple Stain Rat Tissue MAX PO (MULTI) (Nichirei - Code 414191F), for 30 minutes, at room temperature.

      Chromogen: Liquid DAB (Spring - Code DAB-125), for 10 minutes, at room temperature.

      But, unfortunately, the results weren't good. In this test we could see better the immunostaining, but accordingly to our costumer it's non-specific. I mean, the antibody is staining all the tissue structures, being impossible to identify the bone cells (see attached images 1 and 2). At this test reaction, we also did negative control slides (omitting primary antibody) and they are all OK (none immunostaining observed on these slides), what indicates that the non-specific staining observed on the positive slides (incubated with ab52128) are really caused by the primary antibody.

      Our costumer is comparing this new test results with ab52128 with the results obtained with another antibody anti-Bone Sialoprotein (from another manufacturer) she had used before (see attached images 3 and 4). With this other antibody we can see that some structures on the tissue are stained and others no as expected, but the same wasn't observed for ab52128 (compare with images 1 and 2).

      What do you think about that case?

      Do you have any other suggestion for trying to improve protocol?

      I'm forward to hearing from you

      Thanks for your attention.

      Read More
      Answer

      Thank you very much for your reply.

      I am sorry to hear that the results are still not satisfactory, and I appreciate how much time the customer has spenttroubleshooting. It sounds like the antibody is not work as expected, and I would be happy to send a replacement or issue a credit or refund if the customer would prefer. We have a different bone sialoprotein antibody that is guaranteed to work with rat samples in IHC-P, which I can send as a free of charge replacement to the customer-

      https://www.abcam.com/Bone-Sialoprotein-antibody-ab125227.html

      Please let me know if the customer would like to accept one of these options, and I will arrange it for you promptly. I look forward to hearing from you, and if there is anything else that we can do please let me know.

      Read More

      Question

      Our costumer has used Microwave Oven for ab52128 retrieval (Citrate pH 6,0, for 3 cycles of 5 minutes each, full power) on her reactions.

      We are going to do new tests with the antibody accordingly to your instructions.

      For retrieval, I intend to use:agree

      1- Citrate pH 6,0 - Electric Pressure Cooker Riptide - Celerus (125oC, 30 seconds)

      and

      2- Citrate pH 6,0 - Steamer (pre-heated buffer, 95-99oC, for 40 minutes)

      Do you think one of these retrieval protocols can work better for ab52128 than microwave oven?

      What's your suggestion? What kind of retrieval do you recommend?

      I'm forward to hearing from you.

      Thanks a lot for your attention and help.

      Have a nice weekend!

      Read More
      Answer

      Thank you very much for your reply and for sending the additional information.

      We have not tried the microwave method with this antibody, but one of the other methods (steamer or pressure cooker) may work better. I would suggest using the steamer, but only heating the slides for 20-30 minutes to avoid over-retrieving the tissue. The slides can be allowed to cool for 10-20 minutes on the benchtop.

      Please keep me updated about the results after trying the alterations that we've discussed, and if the staining does not improve then I'd be happy to send a replacement or issue a credit or refund. I look forward to hearing from you.

      Have a nice day!

      Read More

      Question

      Dear Mr. or Ms.,

      One of our costumers is having problems with:

      Rabbit Policlonal anti-Bone Sialoprotein

      ABCAM - Code ab52128

      She is using IHC-P technique, rat tissue, and says that the antibody isn't working properly.

      Accordingly to her, it is not giving specific staining in all tested samples (see attached images)

      Tested Protocols with ab52128:

      Samples:

      Rat Mandibular Condyle Bone

      Decalcification: EDTA for 3 months

      Fixation: 10% Formalin for 48 hours

      Tissue Sections: 3 µ

      Tested Retrieval Methods:

      Heating - Citrate, pH 6,0

      Enzymatic - Pepsin and Proteinase K

      Blocking Steps:

      Peroxidase (H2O2) and Protein (BSA, for 1 hour)

      Primary Antibody (ab52128)

      Tested Dilutions: 1:100, 1:300, 1:500 and 1:1000 (at 1:00 and 1:300 there's intense background and at 1:500 and 1:1000 there's less background but no specific staining)

      Incubation: 1 hour, at room temperature

      What do you think about that case? Could it be a problem with antibody vial?

      If you think that the antibody is ok, please, can you give any us any clue to improve protocol?

      I have no direct experience with bone decalcification for IHC uses but aren't 3 months in EDTA too much for the samples?

      I'm forward to hearing from you.

      Thanks for your attention.

      Best Regards,

      Read More
      Answer

      Thank you very much for contacting us and letting us know about the customer's trouble with this antibody.

      Threemonths in EDTA is not too long for decalcification. Bone can be a difficult tissue to work with, soit's possible that there is a problem with thevial but there may also be some protocol alterations that will help.

      I've been in touch with the lab regarding this antibody, and we may have a few recommendations that might improve the results. How long was the tissue heated in the citrate buffer for antigen retrieval?

      For blocking, we would recommend using 5-10% serum to reduce non-specific binding. An overnight incubation with the primary antibody at a higher dilution (1:500 or 1:1000) may also increase the specific binding of the antibody.

      We do fully guarantee this antibody to work in IHC-P with rat tissue, so if the customer is not satisfied with it then we can send a replacement, or issue a credit. Please let me know if the customer would prefer one of these options.

      I look forward to hearing from you. If there is anything else that we can do for you, please let me know and I'll be happy to help.

      Read More

      Question

      Please kindly let me know working dilution possible (WB application) for ab52128 and ab78278 for try. Best regards

      Read More
      Answer

      Thank you for your inquiry. Unfortunatly we do not have a specific dilution to recommend for these antibodies. I can suggest to try a range of dilutions from 1:100 to 1:3000 and to optimize the dilution according to the results. Too much antibody will result in non-specific bands. 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

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