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Anti-DRAM抗体[M3-P4B4] (ab64739)

Reviews (2)Q&A (2)References (1)
Western blot - Anti-DRAM antibody [M3-P4B4] (ab64739)

    Key features and details

    • Mouse monoclonal [M3-P4B4] to DRAM
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG1

    概述

    • 产品名称

      Anti-DRAM抗体[M3-P4B4]
      参阅全部 DRAM 一抗

    • 描述

      小鼠单克隆抗体[M3-P4B4] to DRAM

    • 宿主

      Mouse

    • 经测试应用

      适用于: WBmore details
      不适用于: ICC/IF

    • 种属反应性

      与反应: Human

    • 免疫原

      Synthetic peptide corresponding to Human DRAM aa 150-250.

    • 阳性对照

      • This antibody gave a positive signal in HeLa whole cell lysate.

    • 常规说明

      This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

      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ab64739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 (1)
    Use a concentration of 10 µg/ml. Predicted molecular weight: 26 kDa.
    说明
    WB
    Use a concentration of 10 µg/ml. Predicted molecular weight: 26 kDa.
    应用说明
    Is unsuitable for ICC/IF.

    靶标

    • 功能

      Lysosomal modulator of autophagy that plays a central role in p53/TP53-mediated apoptosis. Not involved in p73/TP73-mediated autophagy.

    • 序列相似性

      Belongs to the DRAM/TMEM150 family.

    • 细胞定位

      Lysosome membrane.
    • Information by UniProt

    • 数据库链接

      • 别名

        • Damage regulated autophagy modulator antibody
        • Damage-regulated autophagy modulator 1 antibody
        • Damage-regulated autophagy modulator antibody
        • DNA damage-regulated autophagy modulator protein 1 antibody
        • DRAM antibody
        • dram1 antibody
        • DRAM1_HUMAN antibody
        • FLJ11259 antibody
        see all

      图片

      • Western blot - Anti-DRAM antibody [M3-P4B4] (ab64739)
        Western blot - Anti-DRAM antibody [M3-P4B4] (ab64739)
        Anti-DRAM antibody [M3-P4B4] (ab64739) at 10 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg

        Secondary
        Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

        Performed under reducing conditions.

        Predicted band size: 26 kDa
        Observed band size: 26 kDa


        Exposure time: 20 minutes

      文献 (1)

      发表研究结果有使用 ab64739？请让我们知道，以便我们可以引用本数据表中的参考文章。

      ab64739 被引用在 1 文献中.

      • Zhang Y  et al. FUT2 Facilitates Autophagy and Suppresses Apoptosis via p53 and JNK Signaling in Lung Adenocarcinoma Cells. Cells 11:N/A (2022). PubMed: 36552800

      客户评价及客户问答

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      1-4 of 4 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-DRAM antibody [M3-P4B4]

       Inconclusive
      Abreviews
      This product is known to not work in this application or species.
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (glioma stem cells)
      Specification
      glioma stem cells
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - triton-x 0.1%
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°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 Apr 26 2012

      Western blot abreview for Anti-DRAM antibody [M3-P4B4]

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Tissue lysate - whole (HeLa Ms Stomach Ms Kidney Ms Liver)
      Loading amount
      20 µg
      Specification
      HeLa Ms Stomach Ms Kidney Ms Liver
      Gel Running Conditions
      Reduced Denaturing
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°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.

      Mr. Antibody Solutions

      Verified customer

      提交于 May 25 2010

      Question

      I am sending you the answer to your questions

      - What is the purchase order number of the product and how it was stored?

      The purchase order number is 23//05/12.It was stored at-20º Cupon arrival after several aliquots were made, stored at -20º C as well.

      - What is the lysate type and species and how it was prepared?

      Each lysate was prepared from 500 000 cells (MM.1S cells, human multiple myeloma cells, growing in suspension). We used SET buffef containing SDS, EDTA and TRIS. Samples were boiled for 15 minutes.

      - Have you added protease inhibitors?

      No

      Was the lysate ice cold?

      After boiling, lysates were kept in ice until loading.

      - The Santa Cruz antibody also shows a large array of bands and no bands with Abcam ab, might be indicating the lysate problem. Have you used similar lysate preparation procedure successfully before?

      We have analyzed the same lysates using an antibody from Sigma. We observed a band at around 27 KDa that was more intense in lysates from apoptotic cells. I am sending you attached a file with these results.

      - How long the lysates were heated in sample buffer?

      15 minutes

      - Could you provide the species of secondary antibody?

      Goat anti-mouse.

      Thank you for your assistance.

      Read More
      Answer

      Thank you very much for confirming the details.

      I have checked the history of this antibody and can confirm that we haven't received specificity complaints for this product so I am sure protocol troubleshooting will help in getting a clean western blot results. I have following suggestions;

      - - In our own lab we test both milk and BSA as blocking solution. 5% BSA, 2 hours incubation time is the most common method and gives excellent results
      - We heat the lysates for 10 minutes at 100C
      - No primary control; this helps checking the non specificity due to secondary antibody
      - The problem might be with the lysates preparation, I would recommend using NP-40 or nonidet-P40 buffer for cell lysis. The ingredients can be found by clicking the following link;

      https://www.abcam.com/index.html?pageconfig=resource&rid=11414

      - For best results we recommend customers adding protease inhibitors. If possible, please add these in lysis buffer.
      - Positive control; Hela cells is the best positive control to be used with this antibody

      I hope the antibody would work following these suggestions. If in case the antibody fails again please contact me again for further assistance.

      Read More

      Question

      Name: Isabel Marzo
      Product code: 64739
      Lot number: 799509
      Inquiry: No specific bands at the reported MW for DRAM were detected. (See image). * Amount of protein loaded: 30 micrograms/lane * Blocking conditions 1 h at r.t. with 5% skimmed milk * Detection method: chemioluminiscence (Pierce) * Electrophoresis and transfer conditions: 12% SDS-PAGE. Semidry transfer. * Primary antibody conditions (diluent/dilution, incubation time/temperature) 1/500 in 2 %(w/v) skimmed milk, 24 h at 4 degrees. * Secondary antibody conditions (diluent/dilution, incubation time/temperature) 1/1000 in 2 %(w/v) skimmed milk, 90 min at r.t. * Wash steps: three washes between blockage-1st antibody, 1st antibody-2nd antibody and before dectection. Buffer containing 0.1% Tween 20.

      Read More
      Answer

      Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

      I have read the details you have kindly provided and would like to ask few questions in order to understand the problem better;

      - What is the purchase order number of the product and how it was stored?
      - What is the lysate type and species and how it was prepared?
      - Have you added protease inhibitors? Was the lysate ice cold?
      - The Santa Cruz antibody also shows a large array of bands and no bands with Abcam ab, might be indicating the lysate problem. Have you used similar lysate preparation procedure successfully before?
      - How long the lysates were heated in sample buffer?
      - Could you provide the species of secondary antibody?

      I am looking forward to hearing from you soon.

      Read More

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