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Cell Biology Apoptosis Extracellular Signals Granzymes

Anti-Granzyme K抗体[GM-24C3] (ab3771)

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Flow Cytometry - Anti-Granzyme K antibody [GM-24C3] (ab3771)

    Key features and details

    • Mouse monoclonal [GM-24C3] to Granzyme K
    • Suitable for: ELISA, Flow Cyt (Intra), Flow Cyt
    • Reacts with: Recombinant fragment
    • Isotype: IgG2b

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    概述

    • 产品名称

      Anti-Granzyme K抗体[GM-24C3]
      参阅全部 Granzyme K 一抗
    • 描述

      小鼠单克隆抗体[GM-24C3] to Granzyme K
    • 宿主

      Mouse
    • 特异性

      This antibody recognises Granzyme K transiently expressed on the cell surface of transfected BOSC cells as well as the native protein in peripheral blood mononuclear cells. It does not cross react with Granzyme A. Specificity is routinely tested by flow cytometry on BOSC cells transiently transfected with a Granzyme K expression vector.
    • 经测试应用

      适用于: ELISA, Flow Cyt (Intra), Flow Cytmore details
    • 种属反应性

      与反应: Recombinant fragment
    • 免疫原

      Other Immunogen Type corresponding to Human Granzyme K. Genetic immunization with cDNA encoding human Granzyme K
      Database link: P49863

    • 阳性对照

      • Flow Cyt: Granzyme K transfected BOSC23 cells.
    • 常规说明


      Granzymes are exogenous serine proteases that are stored in the cytotoxic granules of activated T cells and NK cells. Upon target cell contact, the contents of these granules are directionally exocytosed and, with the assistance of perforin, the granzymes enter the cytosol of the target cell. To date, five human granzymes (A, B, H, K,M) have been described at the molecular genetic level. Human granzyme K (GZMK) is a 28 kD aserine protease whose gene is located on chromosome 5q11-12 close to the granzyme A-encoding gene. Like granzyme A, it has a trypsin-like specifity cleaving at the basic residues arginine and lysine. To which extent human granzyme K plays a role in the induction of apoptosis in the target cells remains to be evaluated. However, granzyme K purified from a rat large granular lymphoma cell line (RNK-16)has been shown to induce apoptosis in vitro. High mRNA levels of granzyme K are detected inactivated T cells and NK cells but are absent in normal tissues that do not contain high numbers of these cells. Antibodies produced from cDNA: Conventional technologies usually either generate antibodies against purified proteins, or against synthetic peptides based on amino acid sequences derived from DNA sequence data. Genetic immunization involves introducing the gene in the form of a cDNA directly into an animal which translates this cDNA into protein thus stimulating an immune response against the foreign protein. Although the synthetic peptide approach is comparable in speed, the quality of antibodies generated by genetic immunization is far superior. This is because the protein is made by the immunized animal, utilzing complex cellular mechanisms that allow it to gain a native conformation. Antibodies are then generated against a native protein, such as is found in the blood or tissues of its host species. Membrane-bound or secreted proteins often create problems for conventional antibody technology because in their native form, they are often modified by glycosylation, or in some cases exist as multiple membrane-spanning proteins that are not soluble following isolation or synthesis in recombinant systems. All of these problems are avoided if the immunized animal makes the protein itself. Antibodies generated by genetic immunization have been shown to have binding affinities to the protein in the sub-nanomolar range, which are approximately 100x higher than conventionally developed antibodies and much higher than single chain antibodies. Results confirm published data for much higher avidity of sera generated by genetic immunization as compared with that gained by immunization with a corresponding recombinant protein.

      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

    • 形式

      Liquid
    • 存放说明

      Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
    • 存储溶液

      pH: 7.20
      Constituent: PBS
    • Concentration information loading...
    • 纯度

      Protein G purified
    • Primary antibody说明

      Granzymes are exogenous serine proteases that are stored in the cytotoxic granules of activated T cells and NK cells. Upon target cell contact, the contents of these granules are directionally exocytosed and, with the assistance of perforin, the granzymes enter the cytosol of the target cell. To date, five human granzymes (A, B, H, K,M) have been described at the molecular genetic level. Human granzyme K (GZMK) is a 28 kD aserine protease whose gene is located on chromosome 5q11-12 close to the granzyme A-encoding gene. Like granzyme A, it has a trypsin-like specifity cleaving at the basic residues arginine and lysine. To which extent human granzyme K plays a role in the induction of apoptosis in the target cells remains to be evaluated. However, granzyme K purified from a rat large granular lymphoma cell line (RNK-16)has been shown to induce apoptosis in vitro. High mRNA levels of granzyme K are detected inactivated T cells and NK cells but are absent in normal tissues that do not contain high numbers of these cells. Antibodies produced from cDNA: Conventional technologies usually either generate antibodies against purified proteins, or against synthetic peptides based on amino acid sequences derived from DNA sequence data. Genetic immunization involves introducing the gene in the form of a cDNA directly into an animal which translates this cDNA into protein thus stimulating an immune response against the foreign protein. Although the synthetic peptide approach is comparable in speed, the quality of antibodies generated by genetic immunization is far superior. This is because the protein is made by the immunized animal, utilzing complex cellular mechanisms that allow it to gain a native conformation. Antibodies are then generated against a native protein, such as is found in the blood or tissues of its host species. Membrane-bound or secreted proteins often create problems for conventional antibody technology because in their native form, they are often modified by glycosylation, or in some cases exist as multiple membrane-spanning proteins that are not soluble following isolation or synthesis in recombinant systems. All of these problems are avoided if the immunized animal makes the protein itself. Antibodies generated by genetic immunization have been shown to have binding affinities to the protein in the sub-nanomolar range, which are approximately 100x higher than conventionally developed antibodies and much higher than single chain antibodies. Results confirm published data for much higher avidity of sera generated by genetic immunization as compared with that gained by immunization with a corresponding recombinant protein.
    • 克隆

      单克隆
    • 克隆编号

      GM-24C3
    • 同种型

      IgG2b
    • 研究领域

      • Cell Biology
      • Apoptosis
      • Extracellular Signals
      • Granzymes
      • Immunology
      • Adaptive Immunity
      • T Cells
      • Cytotoxic Cells
      • Immunology
      • Innate Immunity
      • NK Cells

    相关产品

    • Compatible Secondaries

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Conjugation kits

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

      • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
    • Recombinant Protein

      • Recombinant human Granzyme K protein (ab157277)

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab3771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
    ELISA
    Use at an assay dependent concentration.
    Flow Cyt (Intra)
    Use at an assay dependent concentration.

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

    Flow Cyt
    Use 1.2µg for 106 cells.
    说明
    ELISA
    Use at an assay dependent concentration.
    Flow Cyt (Intra)
    Use at an assay dependent concentration.

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

    Flow Cyt
    Use 1.2µg for 106 cells.

    靶标

    • 组织特异性

      Expressed in lung, spleen, thymus and peripheral blood leukocytes.
    • 序列相似性

      Belongs to the peptidase S1 family. Granzyme subfamily.
      Contains 1 peptidase S1 domain.
    • 细胞定位

      Secreted. Cytoplasmic granule.
    • Target information above from: UniProt accession P49863 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • 别名

      • Fragmentin 3 antibody
      • Fragmentin-3 antibody
      • Fragmentin3 antibody
      • GRAK_HUMAN antibody
      • Granzyme 3 antibody
      • granzyme K (granzyme 3; tryptase II) antibody
      • granzyme K (serine protease, granzyme 3; tryptase II) antibody
      • Granzyme K antibody
      • Granzyme K precursor antibody
      • Granzyme-3 antibody
      • Granzyme3 antibody
      • GranzymeK antibody
      • GZMK antibody
      • NK TRYP 2 antibody
      • NK TRYP2 antibody
      • NK tryptase 2 antibody
      • NK-TRYP-2 antibody
      • NK-tryptase-2 antibody
      • NKTRYP2 antibody
      • Serine protease granzyme 3 antibody
      • TRYP 2 antibody
      • TRYP2 antibody
      • Tryptase II antibody
      • TryptaseII antibody
      see all

    图片

    • Flow Cytometry - Anti-Granzyme K antibody [GM-24C3] (ab3771)
      Flow Cytometry - Anti-Granzyme K antibody [GM-24C3] (ab3771)

      Flow cytometric analysis of BOSC23 cells using ab3771. BOSC23 cells were transiently transfected with an expression vector encoding either Granzyme K (red curve) or an irrelevant protein (control transfectant: black curve). Binding of ab3771 was detected with a PE-conjugated secondary antibody. A positive signal was obtained only with Granzyme K transfected cells.

    实验方案

    • Flow cytometry protocols

    Click here to view the general protocols

    数据表及文件

    • Datasheet download

      Download

    文献 (0)

    发表研究结果有使用 ab3771?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab3771 尚未被引用在任何文献中。

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-2 of 2 Abreviews or Q&A

    Question

    I have recently bought 2 antibodies from Abcam. There are polyclonal Granzyme H (cat#ab58852) and monoclonal Granzyme K (cat#ab3771) for IHC purposes. May I know what is the suggested positive and negative tissue controls for these 2 antibodies in IHC. Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 18 2008

    Answer

    Thank you for your enquiry. Unfortunately, there was not much information I could find regarding suitable negative controls for these products, but I did manage to find information on positive controls in Swiss Prot. ab3771 - expression can be found in lung, spleen, thymus and peripheral blood leukocytes. Therefore, you can use lung and spleen as positive controls in IHC. ab58852 - product was tested on human kidney tissue and this would be our suggested positive control. I hope this information helps. If there is anything else that I can help you with, please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    回复于 Nov 18 2008

    Question

    We are now looking for Granzyme K assay kit which I find three from your company webpage (ab3771, ab26150, and ab26155). However, I would like to clarify something first before making final decision: 1. As three items are available for the same antibody, what are the difference between these three items? Do they only different in clones or any other aspect? Please advise. 2. From the webpage, all three items are tested for ELISA, Flow Cytometry / FACS application. However, I would like to employ the antibody for immunohistostaining work on formalin-paraffin block. Is it possible? If yes, any recommended protocols can be provided from your side, and any additional reagents are needed? 3. If I choose it to work in ELISA format, any secondary antibody, conjugate, and substrate, is recommended from your company?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 12 2006

    Answer

    Thank you for your enquiry. The three antibodies against granzyme K (ab3771, ab2615, and ab26155) will likely recognize different epitopes and have different affinities for the protein however this has not been confirmed. All three have been tested for the use in flow cytometry and ELISA but not in immunohistochemistry, so we do not know if they will work in this application. For flow cytometry, ab3771 achieved the brightest signals. For development of an ELISA for soluble GzmK, the antibody combination ab26150 as the catcher Ab and biotinylated ab26155 as the detector Ab achieved worked best. Although many different types of enzymes have been used for detection, horse radish peroxidase (HRP) and alkaline phosphatase (ALP) are the two widely used enzymes used in ELISA assay. I hope this information helps. Please contact me if you have further questions.

    Read More

    Abcam Scientific Support

    回复于 Jul 13 2006

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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