Anti-PERP antibody (ab5986)

Reviews (1)Q&A (5)References (6)

Overview

  • Product name

    Anti-PERP antibody
    See all PERP primary antibodies

  • Description

    Rabbit polyclonal to PERP

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-Fr, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse

  • Immunogen

    Synthetic peptide:

    EDDLLGNAKPRYFYTSA

    , corresponding to amino acids 175-193 of Human PERP.

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

  • Positive control

    • A431 whole cell lysate.

  • General notes

    Stable for one year.


    The p53 tumor-suppressor gene integrates numerous signals that control cell life and death. Several novel molecules involved in the p53 network, including Chk2, p53R2, p53AIP1, Noxa, PIDD, PID/MTA2, MTBP and PERP, were identified and their genes were cloned recently. PERP, also termed PIGPC1 and THW, is a plasma membrane protein. p53 binds to the promoter of PERP and transcriptionally activates PERP gene then the translated PERP protein mediates the p53 induced apoptosis. The expression of PERP causes cell death. PERP is a mediator of p53 induced apoptosis. PERP has sequence similarity to PMP-22/gas3 and is a new member of the PMP-22/gas3 family.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Primary antibody notes

    The p53 tumor-suppressor gene integrates numerous signals that control cell life and death. Several novel molecules involved in the p53 network, including Chk2, p53R2, p53AIP1, Noxa, PIDD, PID/MTA2, MTBP and PERP, were identified and their genes were cloned recently. PERP, also termed PIGPC1 and THW, is a plasma membrane protein. p53 binds to the promoter of PERP and transcriptionally activates PERP gene then the translated PERP protein mediates the p53 induced apoptosis. The expression of PERP causes cell death. PERP is a mediator of p53 induced apoptosis. PERP has sequence similarity to PMP-22/gas3 and is a new member of the PMP-22/gas3 family.

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5986 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 10 µg/ml.
IHC-Fr Use at an assay dependent concentration. PubMed: 18989386
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 21 kDa.

Target

  • Function

    Component of intercellular desmosome junctions. Plays a role in stratified epithelial integrity and cell-cell adhesion by promoting desmosome assembly. Plays a role as an effector in the TP53-dependent apoptotic pathway.

  • Tissue specificity

    Expressed in skin, heart, placental, liver, pancreas, keratinocytes and dermal fibroblasts.

  • Sequence similarities

    Belongs to the TMEM47 family.

  • Cellular localization

    Cell junction > desmosome. Cell membrane. Associated with desmosomes.
  • Information by UniProt

  • Database links

    • Alternative names

      • 1110017A08Rik antibody
      • dJ496H19.1 antibody
      • KCP 1 antibody
      • KCP-1 antibody
      • KCP1 antibody
      • Keratinocyte associated protein 1 antibody
      • Keratinocyte-associated protein 1 antibody
      • Keratinocytes associated protein 1 antibody
      • KRTCAP 1 antibody
      • KRTCAP1 antibody
      • p53 apoptosis effector related to PMP 22 antibody
      • p53 apoptosis effector related to PMP-22 antibody
      • p53 apoptosis effector related to PMP22 antibody
      • P53 induced protein PIGPC1 antibody
      • P53-induced protein PIGPC1 antibody
      • Perp antibody
      • PERP TP53 apoptosis effector antibody
      • PERP_HUMAN antibody
      • PIGPC 1 antibody
      • PIGPC1 antibody
      • RP3 496H19.1 antibody
      • THW antibody
      • TP53 apoptosis effector antibody
      • Transmembrane protein THW antibody
      see all

    Images

    • Western blot - Anti-PERP antibody (ab5986)
      Western blot - Anti-PERP antibody (ab5986)
      Western blot analysis of PERP expression in A431 whole cell lysates in the absence (A) and presence (B) of blocking peptide with anti- PERP at 1 µg /ml. Western blot analysis of PERP expression in A431 whole cell lysates in the absence (A) and presence (B) of blocking peptide with anti- PERP at 1 µg /ml.
    • Immunocytochemistry/ Immunofluorescence - Anti-PERP antibody (ab5986)
      Immunocytochemistry/ Immunofluorescence - Anti-PERP antibody (ab5986)
      Immunocytochemistry of PERP in A431 cells with PERP antibody at 10 µg/ml.

    References

    This product has been referenced in:

    • Awais R  et al. p63 is required beside p53 for PERP-mediated apoptosis in uveal melanoma. Br J Cancer 115:983-992 (2016). WB . Read more (PubMed: 27584665) »
    • Marthandan S  et al. Similarities in Gene Expression Profiles during In Vitro Aging of Primary Human Embryonic Lung and Foreskin Fibroblasts. Biomed Res Int 2015:731938 (2015). WB . Read more (PubMed: 26339636) »
    See all 6 Publications for this product

    Publishing research using ab5986? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-5 of 5 Q&A

    Question


    I called earlier to inquire about this product and decided to exchange this order for ab5986 upon hearing that ab48032 would be backordered for awhile. I have tested out ab5986, which is only predicted to work on mouse, on mouse mammary tumors for IF and obtained negative results with two different processing methods; these methods worked perfectly well for ab3945, which I believe has been discontinued by your company. Would I still be able to use the discount code to try out ab48032 on mouse tissue? Thank you.
    Best,

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    Answer

    Thank you for your email and for keeping me updated about the situation.
    I'm sorry to hear that ab5986 did not work well with the mouse samples, and it is possible that this antibody just doesn't work with mouse. We have unfortunately removed ab3945 from our catalog, but the discount code for ab48032 is still valid if you would like to try that antibody. The code expires in August, but if you need to extend that date please let me know and I'll be happy to issue you a new code.
    Please let me know if you have any further questions or if there is anything else that we can do for you.

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    Question

    Would like to try ab48032 with mouse samples in ICC/IF.

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    Answer

    Thank you for your call today and for your interest in ab48032. Please see below for the details of your testing discount code, and let me know if you have any questions or if there is anything else that we can do for you.
    DISCOUNT CODE: ***
    Expiration date: August 24, 2012
    This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab48032 using mouse samples and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
    The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.
    The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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    Question

    1. Order details: • Batch number: SA1217-AB5986 • Abcam order or Purchase order number: B09501149 • Antibody storage conditions (temperature/reconstitution etc) ????10????,???4???;????????? 2. Please describe the problem (high background, wrong band size, more bands, no band etc). no band 3. On what material are you testing the antibody in WB? • Species: cell line • Cell extract or Nuclear extract: Cell extract • Purified protein or Recombinant protein: Recombinant protein 3. The lysate • How much protein was loaded: 30?g • What lysis buffer was used: RIPA • What protease inhibitors were used: co cktail(Rocho) • What loading buffer was used: 2x sample buffer • Did you heat the samples: temperature and time: Yes,95? for 5 min. 4. Electrophoresis/Gel conditions/ Transfer conditions • Reducing or non reducing gel: I don`t understand what you mean. • Gel percentage : stacking: 5?; separating:12.5? • Transfer conditions: 70V for 40min 5. Blocking conditions • Buffer: 5?MILK(in TTBS) • Blocking agent: milk, BSA, serum, what percentage: 5?MILK(in TTBS) • Incubation time: 1hr • Incubation temperature: Room Temperature 6. Primary Antibody • Specification (in which species was it raised against): on ice • At what dilution(s) have you tested this antibody: 1:1000 • What dilution buffer was used: 5?MILK(in TTBS) • Incubation time: O/N • Incubation temperature: 4? • What washing steps were done: TTBS 15min/?X4 7. Secondary Antibody • Specification (in which species was it raised against)? on ice • At what dilution(s) have you tested this antibody: 1:1000 • Incubation time 1hr • Wash steps: TTBS 15min/?X4 • Do you know whether the problems you are experiencing come from the secondary? no 8. Detection method ECl, ECl+, other detection method: ECl 9. Background bands • Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): No, I haven`t. • Is the blocking step sufficient? Yes. • Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) Yes • At what size are the bands migrating? Could they be degradation products of your target? Between17-24 kd • Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) • ? 10min ? 1hr 11. Did you apply positive and negative controls along with the samples? Please specify. No 10. Optimization attempts • How many times have you tried the Western? 1 • Do you obtain the same results every time e.g. are background bands always in the same place? • What steps have you altered? Please send this questionnaire by e-mail by cutting and pasting the text in your e-mail (no attachements other than images) to technical@abcam.com.

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    Answer

    Thank you for the details that you have provided and I'm sorry to hear that you are experiencing difficulty with ab5986. At this point I would like to make the following suggestions to help improve results as you are not seeing any bands. This antibody was characterized for use in Western blotting with A431 lysate and I strongly suggest using this as a positive control. I also suggest increasing the amount of primary that you are using to a dilution of 1:500 and incubating overnight at 4C. Also, you want to ensure that the protein transferred properly to the membrane (check this with Ponceau S) and check that your secondary is working properly. Is the secondary working with other primaries? You may also need to increase the incubation period and concentration of your secondary. I hope this helps. Please contact us again if you need additional assistance.

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    Question

    BATCH NUMBER 76164 ORDER NUMBER 61814 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE Cell extracts PRIMARY ANTIBODY ab5986 was the primary antibody. Different dilutions were used form 1:500 to 1:5000. Antibody was diluted in TBS-Tween 5% milk. Incubation time: 1-1h30min Wash step: One rapid was and 3x10 min washes witn TBS-Tween. SECONDARY ANTIBODY Anti-rabbit IgG HRP (Amersham NA934V)was the secondary antibody. Different dilutions were used form 1:20000 to 1:50000. Antibody was diluted in TBS-Tween 5% milk. Incubation time: 1-1h30min Wash step: One rapid was and 3x10 min washes witn TBS-Tween. DETECTION METHOD Pierce Super Signal West Pico POSITIVE AND NEGATIVE CONTROLS USED Yes. Positive control: A431 lysate ab7909. Negative control: cell extract from bacteria. ANTIBODY STORAGE CONDITIONS Stored at +4?C SAMPLE PREPARATION Laemmli buffer has been used to prepare the samples. An equel volume of 2x Laemmli sample buffer was added. The sample buffer was complemented with b-mercaptoethanol. Samples are heated at 95?C for 5 min before loading them in the gel. AMOUNT OF PROTEIN LOADED 50-100 ng of target protein were loaded. ELECTROPHORESIS/GEL CONDITIONS 12% SDS-PAGE. TRANSFER AND BLOCKING CONDITIONS Tris-Glycine-methanol transfer buffer was used for running the gel. Gels were trasnfered for 45 min at 120V. The efficiency of trasnfer was determined by staining the membrane with Ponceau solution. Blocking reagent used was TBS-Tween buffer with 5% milk. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 15 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

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    Answer

    I'm sorry to hear you are having a problem with ab5986. I would like to suggest an overnight incubation of the antibody at 4C and maybe try a dilution of 1:300 as well. If you still have problems and your positive control (50ug of lysate per well) clearly shows that the antibody does not work please let me know and please send me an image of your blot and I will arrange for a replacement to be sent to you.

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    Question

    As you requested, here is the feedback from the replacement antibody for PERP (ab5986). Please see attached image of WB, obtained with 1 ug/ml of Ab (ab5986), standard conditions, as described in my previous message (regarding use of the other PERP Ab - ab 3945). M=size markers, as indicated; A431 cell lysate, as positive control; 1-4 our samples. Equal total protein loading in all lanes, as demonstrated by GAPDH probing. ab 5986 appears to recognize a band of the expected size, i.e. 21kDa, which was not recognized by the other Ab. However, note that two other bands appear, with the 37 kDa one being the band which appeared strongly immunopositive with the ab 3945, as described previously. Your comments regarding this will be appreciated. Best regards, Luminita.

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    Answer

    Thank you for getting back to us and providing us some more information about your recent results. We can see on the Western blot image that the 21 kDa band appears nicely, however there are two other non-specific bands as well. It is important to emphasize that ab5986 is a polyclonal antibody and it recognizes different epitopes. Therefore the antibody may recognize other proteins as well which share similar amino acid sequences. We would suggest perhaps reducing the concentration of the primary antibody and applying more stringent washings. Have you tested the secondary antibody alone? We hope this will help.

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