Anti-Perforin antibody (ab7203)

Submit a review Q&A (7)References (3)

Overview

  • Product name

    Anti-Perforin antibody
    See all Perforin primary antibodies

  • Description

    Rabbit polyclonal to Perforin

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat

  • Immunogen

    Recombinant full length protein (Rat).

  • Positive control

    • ICC/IF: SW480 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab7203 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
ICC/IF Use a concentration of 5 µg/ml.
WB 1/1000. Predicted molecular weight: 70 kDa.

Target

  • Function

    Plays a key role in secretory granule-dependent cell death, and in defense against virus-infected or neoplastic cells. Plays an important role in killing other cells that are recognized as non-self by the immune system, e.g. in transplant rejection or some forms of autoimmune disease. Can insert into the membrane of target cells in its calcium-bound form, oligomerize and form large pores. Promotes cytolysis and apoptosis of target cells by facilitating the uptake of cytotoxic granzymes.

  • Involvement in disease

    Defects in PRF1 are the cause of hemophagocytic lymphohistiocytosis familial type 2 (FHL2) [MIM:603553]; also known as HPLH2. Familial hemophagocytic lymphohistiocytosis (FHL) is a genetically heterogeneous, rare autosomal recessive disorder. It is characterized by immune dysregulation with hypercytokinemia and defective natural killer cell function. The clinical features of the disease include fever, hepatosplenomegaly, cytopenia, hypertriglyceridemia, hypofibrinogenemia, and neurological abnormalities ranging from irritability and hypotonia to seizures, cranial nerve deficits, and ataxia. Hemophagocytosis is a prominent feature of the disease, and a non-malignant infiltration of macrophages and activated T lymphocytes in lymph nodes, spleen, and other organs is also found.

  • Sequence similarities

    Belongs to the complement C6/C7/C8/C9 family.
    Contains 1 C2 domain.
    Contains 1 EGF-like domain.
    Contains 1 MACPF domain.

  • Domain

    The C2 domain mediates calcium-dependent binding to lipid membranes. A subsequent conformation change leads to membrane insertion of beta-hairpin structures and pore formation. The pore is formed by transmembrane beta-strands.

  • Post-translational
    modifications

    N-glycosylated.

  • Cellular localization

    Cytoplasmic granule lumen. Secreted. Cell membrane. Endosome lumen. Stored in cytoplasmic granules of cytolytic T-lymphocytes and secreted into the cleft between T-lymphocyte and target cell. Inserts into the cell membrane of target cells and forms pores. Membrane insertion and pore formation requires a major conformation change. May be taken up via endocytosis involving clathrin-coated vesicles and accumulate in a first time in large early endosomes.
  • Information by UniProt

  • Database links

    • Alternative names

      • Cytolysin antibody
      • FLH2 antibody
      • HPLH2 antibody
      • Lymphocyte pore-forming protein antibody
      • P1 antibody
      • PERF_HUMAN antibody
      • perforin 1 (pore forming protein) antibody
      • Perforin 1 antibody
      • Perforin-1 antibody
      • PFP antibody
      • PGFL antibody
      • PIGF antibody
      • PIGF-2 antibody
      • PLGF antibody
      • Pore forming protein antibody
      • prf1 antibody
      • SHGC-10760 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-Perforin antibody (ab7203)
      Immunocytochemistry/ Immunofluorescence - Anti-Perforin antibody (ab7203)

      ICC/IF image of ab7203 stained SW480 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7203, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Western blot - Anti-Perforin antibody (ab7203)
      Western blot - Anti-Perforin antibody (ab7203)
      Western blot detection of rat Perforin from kidney on Day 3 in a glomerular nephritis model.

    References

    This product has been referenced in:

    • Rothhammer V  et al. a4-integrins control viral meningoencephalitis through differential recruitment of T helper cell subsets. Acta Neuropathol Commun 2:27 (2014). WB . Read more (PubMed: 24606807) »
    • Kish DD  et al. Neutrophil expression of Fas ligand and perforin directs effector CD8 T cell infiltration into antigen-challenged skin. J Immunol 189:2191-202 (2012). WB . Read more (PubMed: 22815291) »
    See all 3 Publications for this product

    Publishing research using ab7203? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-7 of 7 Abreviews or Q&A

    Question

    Hello

    I have performed Western Blot with this antibody at the manufacturer’s recommended dilution. However, the antibody is not working properly. I am getting nonspecific bands all over the place. Is it possible to obtain a positive control from you? This way I may be able to titrate this antibody and get publishable results.

    Thanks

    Read More
    Answer

    Thank you for contacting us and your patience, as we were closed over Easter.
    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.
    As positive control, I can either recommend either fresh PBL or inflamed tissues, as well as the YT-Indy cell line - see either SwissProt or our datasheet of the ab86319 for reference: http://www.uniprot.org/uniprot/P10820 https://www.abcam.com/index.html?datasheet=86319
    We normally do not send out positive controls for our antibodies. However we do guarantee all our products to work as stated on the datasheet. I would therefore suggest that we firstly investigate your results.
    In order to do so, 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

    We spoke over the telephone when I advised that I was using this antibody in WB and requested more information about a protocol. In particular, I am interested in how the cell lysate was prepared and whether the blot was performed under reducing or non-reducing conditions. Results so far show many background bands in addition to the expect band at 66kDa, but similar results with the -ve control lysate.

    Read More
    Answer

    We passed your enquiry on to the originator of this antibody and they have replied with the following response. If you have any further questions, please let us know. Many rat kidney models are known to contain anti-GBM antibody, which has approximately 20% cross-reactivity with anti-rabbit IgG. Since anti-rabbit IgG is the secondary Ab used to detect this rabbit polyclonal to perforin, there would be background if the sample was not well-denatured. A complete denaturing of the sample should improve the specificity.

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    Question

    I want to use anti-perforin(CAT ab7203) to block the cytotoxic activities of perforin by abdomal injection. Did you perform similar experiment with this reagent? And how much dose could you please give some advice? I am looking forward to hearing from you soon. Thank you very much.

    Read More
    Answer

    Thank you for your enquiry and yiur interest in our products. Abcam must draw your attention to the disclaimer as set out in its Terms and Conditions: " FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE", and it cannot endorse or otherwise encourage you to use its products for uses other that research. This particular antibody has only been tested for Western blot application.

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    Question

    I know that all of the previous questions have been related to flow cytometry, but the most recent is one year old. Has it been tested for flow cytomtetry in the last year?

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    Answer

    Not to the best of our knowledge.

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    Question

    Need to use antibody to assess expression of Perforin in Human cells by flowcytometry/ by ELISA in culture supernates.

    Read More
    Answer

    The anti-rat perforin has not been tested for use in flow cytometry. Cross reactivity with other species has also not been determened.

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    Question

    I am interested in the rabbit polyclonal antibody to rat/mouse perforin (antibody name 7203). The information that I was able to obtain from your website stated that the application for this antibody was for Western blotting. I was curious to know if it has been tested for use in flow cytometry, and if so, does it work well? I am also interested in getting the most current pricing information. Thank you for your help.

    Read More
    Answer

    This antibody has not yet been tested for use in flow cytometry.

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    Question

    Can the antibody be used for flow cytometry to test for the presence of perfornin in mouse T cells?

    Read More
    Answer

    This antibody has not been tested for use in flow cytometry.

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    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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