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Anti-Cathepsin K抗体[3F9] (ab37259)

Submit a review Q&A (2)References (32)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin K antibody [3F9] (ab37259)
  • Western blot - Anti-Cathepsin K antibody [3F9] (ab37259)
  • Flow Cytometry - Anti-Cathepsin K antibody [3F9] (ab37259)

Key features and details

  • Mouse monoclonal [3F9] to Cathepsin K
  • Suitable for: WB, IHC-P, Flow Cyt
  • Reacts with: Mouse, Human
  • Isotype: IgG2b

概述

  • 产品名称

    Anti-Cathepsin K抗体[3F9]
    参阅全部 Cathepsin K 一抗

  • 描述

    小鼠单克隆抗体[3F9] to Cathepsin K

  • 宿主

    Mouse

  • 经测试应用

    适用于: WB, IHC-P, Flow Cytmore details

  • 种属反应性

    与反应: Mouse, Human

  • 免疫原

    Recombinant fusion protein, corresponding to amino acids 115-329 of Human Cathepsin K. The protein contains at the N-terninal end a His tag (6x) and 6 additional amino acid residues (MRGSHHHHHHGS).

  • 表位

    The monoclonal antibody ab37259 (clone 3F9) reacts with an epitope located between aa 115-329: APDSVDYRKKGYVTPNQGQCGSCWAFSSVGALEGQLKKKTGKLLNLSPQNLVDCVSENDGC GGGYMTNAFQYVQKNRGIDSEDAYPYVGQEESCMYNPTGKAAKCRGYREIPEGNEKALKRA VARVGPVSVAIDASLTSFQFSKGVYYDESCNSDNLNHAVLAVGYGIQKGNKHWIIKNSWGE NWGNKGYILMARNKNNACGIANLASFPKM

  • 常规说明

    This product was changed from ascites to tissue culture supernatant on 22nd December 2017. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team

    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ab37259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
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37, 40 kDa (predicted molecular weight: 37 kDa).
IHC-P
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. 
说明
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37, 40 kDa (predicted molecular weight: 37 kDa).
IHC-P
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. 

靶标

  • 功能

    Closely involved in osteoclastic bone resorption and may participate partially in the disorder of bone remodeling. Displays potent endoprotease activity against fibrinogen at acid pH. May play an important role in extracellular matrix degradation.

  • 组织特异性

    Predominantly expressed in osteclasts (bones).

  • 疾病相关

    Defects in CTSK are the cause of pycnodysostosis (PKND) [MIM:265800]. PKND is an autosomal recessive osteochondrodysplasia characterized by osteosclerosis and short stature.

  • 序列相似性

    Belongs to the peptidase C1 family.

  • 细胞定位

    Lysosome.
  • Information by UniProt

  • 数据库链接

    • 别名

      • Cathepsin K antibody
      • Cathepsin O antibody
      • Cathepsin O1 antibody
      • Cathepsin O2 antibody
      • Cathepsin X antibody
      • CATK_HUMAN antibody
      • CTS02 antibody
      • Ctsk antibody
      • CTSO antibody
      • CTSO1 antibody
      • CTSO2 antibody
      • MGC23107 antibody
      • PKND antibody
      • PYCD antibody
      see all

    图片

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin K antibody [3F9] (ab37259)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin K antibody [3F9] (ab37259)
      Bone from mouse foot (bone marrow with osteoclasts (positive). IHC-P image was obtained after deparaffinization, antigen retrieval with 0.1 M EDTA (6-8 min, 72 C), drying of slide before staining (10 min, 62 C), and subsequent staining with primary antibody (1/100) in 1% horse serum (1 h, 37 C).
    • Western blot - Anti-Cathepsin K antibody [3F9] (ab37259)
      Western blot - Anti-Cathepsin K antibody [3F9] (ab37259)
      Anti-Cathepsin K antibody [3F9] (ab37259) at 1 µg/ml + Human bone tumor tissue lysate - total protein (ab29359) at 10 µg

      Secondary
      Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 37 kDa
      Observed band size: 37,40 kDa why is the actual band size different from the predicted?
      Additional bands at: 26 kDa (possible cleavage fragment)


      Exposure time: 1 minute


      Cathepsin K contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted, seen at 40 kDa. The band observed at 25 kDa could potentially be a cleaved form of Cathepsin K due to the presence of both a 15 amino acid signal prptide and a 99 amino acid propeptide. Cathepsin K in its mature form has an expected molecular weight of 25 kDa.
    • Flow Cytometry - Anti-Cathepsin K antibody [3F9] (ab37259)
      Flow Cytometry - Anti-Cathepsin K antibody [3F9] (ab37259)
      Overlay histogram showing U20S cells stained with ab37259 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37259, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in U20S cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    文献 (32)

    发表研究结果有使用 ab37259？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab37259 被引用在 32 文献中.

    • Kondo S  et al. Elastin microfibril interface-located protein 1 and its catabolic enzyme, cathepsin K, regulate the age-related structure of elastic fibers in the skin. J Cosmet Dermatol 21:4796-4804 (2022). PubMed: 35064622
    • Zhang W  et al. α-Mangostin inhibits LPS-induced bone resorption by restricting osteoclastogenesis via NF-κB and MAPK signaling. Chin Med 17:34 (2022). PubMed: 35248101
    • Ma Y  et al. P2X7 receptor knockdown suppresses osteoclast differentiation by inhibiting autophagy and Ca2+/calcineurin signaling. Mol Med Rep 25:N/A (2022). PubMed: 35266012
    • Arandjelovic S  et al. ELMO1 signaling is a promoter of osteoclast function and bone loss. Nat Commun 12:4974 (2021). PubMed: 34404802
    • Zhang B  et al. DUSP6 expression is associated with osteoporosis through the regulation of osteoclast differentiation via ERK2/Smad2 signaling. Cell Death Dis 12:825 (2021). PubMed: 34475393
    View all Publications for this product

    客户评价及客户问答

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    1-2 of 2 Abreviews or Q&A

    Question

    Buongiorno,

    vi scrivo in italiano e spero riusciate a tradurre.

    Sto cercando di mettere a punto l’Ab Cathepsin K (ab37259) con il sistema di rivelazione Dako FLEX+. Dal data sheet si evince che il seguente metodo:

    1) antigen retrieval : EDTA 6-8’ a 72°C
    2) passaggio successivo: asciugare le sezioni 10’ a 62°C
    3) incubazione Ab primario ( diluito 1:100 in 1% di siero normale di cavallo ) per 1h a 37°C


    domande:

    1.Perchè l’antigen retrieval solo a 72°C anziché i canonici 98°C?
    2.Perché asciugare le sezioni, dato che è una certezza da sempre non dover mai asciugare le sezioni?
    3.Perché diluire l’Ab primario in horse serum 1% e perché incubarlo a 37°C? E’ forse molto debole? Infatti la diluizione consigliata prevede una concentrazione a 10µg/ml… abbastanza alta


    Resto in attesa di chiarimenti

    Grazie per la collaborazione

    Read More
    Answer

    Grazie per la sua domanda. Vi rispondo in Inglese - se ha bisogno di una traduzione, mi faccia sapere per favore. Grazie!
    Thank you for your inquiry.
    The protocol on the datasheet is this protocol is from a customer who tested our antibody and provided his results and photo. The information about the drying is from him too. Unfortunately we do not know more.
    As the tissue he used was bone as well, I would suspect that on bone tissue you can have special protocols (albeit decalcification is before fixation normally).
    In any case, I can recommend you to follow your protocol which is established in your laboratory. I would recommend a normal heat induced antigen retrieval with boiling and only if your tissue is very sensitive, to do antigen retrieval overnight at 70°C. I would also recommend to test different antigen retrieval solutions at different pHs, as we recommend for all our antibodies.
    I do also absolutely agree with you, that after rehydratation the sections should never dry out again, as this will impair the staining strongly.
    The antibody incubation can be done at 37°C, however we recommend to incubate the antibody either 1 to 2 hours at room temperature or overnight at 4°C. This will be absolutely sufficient.
    So in summary, I can recommend you to perform the immunostaining with this antibody as you would do normally. We do guarantee this antibody to work also with a classical IHC protocol.
    I hope this information is helpful. Please do not hesitate to contact us again should you have any further question.

    Read More

    Question

    Dear Colleagues

    We have a customer that asks us, referred to ab37259, why you recommend the following steps: “Bone from mouse foot (bone marrow with osteoclasts (positive). IHC-P image was obtained after deparaffinization, antigen retrieval with 0.1 M EDTA (6-8 min, 72 C), drying of slide before staining (10 min, 62 C), and subsequent staining with primary antibody (1/100) in 1% horse serum (1 h, 37 C)”

    Thanks a lot for your precious collaboration

    With my best regards

    Read More
    Answer

    Thank you for contacting us and your interest in our products.

    I am sorry if the product datasheet has been misleading for your customer. The details provided with the image are to illustrate how this particular IHC staining experiment was performed. They are not intended as a recommendation of how IHC experiments have to be performed. For example the following reference have not used this method when staining human cancer tissue:

    Birgersdotter A et al. Inflammation and tissue repair markers distinguish the nodular sclerosis and mixed cellularity subtypes of classical Hodgkin's lymphoma. Br J Cancer 101:1393-401 (2009). PubMed: 19773754

    The information is provided as a guideline only and optimization is likely to be required depending on the tissue and protocol used by the customer. More information on how to optimize IHC experiments can be found from the following link:

    https://www.abcam.com/index.html?pageconfig=resource&rid=11384

    I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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