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Epigenetics and Nuclear Signaling DNA / RNA DNA Damage & Repair DNA Damage Response p53

Anti-53BP1抗体(ab21083)

  • Datasheet
  • SDS
Reviews (5)Q&A (6)References (104)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)
  • Western blot - Anti-53BP1 antibody (ab21083)
  • Western blot - Anti-53BP1 antibody (ab21083)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)
  • Western blot - Anti-53BP1 antibody (ab21083)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)

Key features and details

  • Rabbit polyclonal to 53BP1
  • Suitable for: WB, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-53BP1 antibody [EPR2172(2)] (ab175933)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-53BP1抗体
    参阅全部 53BP1 一抗
  • 描述

    兔多克隆抗体to 53BP1
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, IHC-Pmore details
  • 种属反应性

    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide corresponding to Human 53BP1 (C terminal). Conjugated to a carrier protein.
    Database link: Q12888

  • 常规说明

    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 freeze / thaw cycles.
  • 存储溶液

    pH: 7.00
    Preservative: 0.025% Proclin 300
    Constituents: 79% PBS, 20% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • p53
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • BRCT Domain Proteins
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • p53 pathway

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab21083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 (2)
1/500 - 1/3000. Detects a band of approximately 350 kDa (predicted molecular weight: 220 kDa).
IHC-P
1/1000 - 1/4000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
说明
WB
1/500 - 1/3000. Detects a band of approximately 350 kDa (predicted molecular weight: 220 kDa).
IHC-P
1/1000 - 1/4000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能

    May have a role in checkpoint signaling during mitosis. Enhances TP53-mediated transcriptional activation. Plays a role in the response to DNA damage.
  • 疾病相关

    Note=A chromosomal aberration involving TP53BP1 is found in a form of myeloproliferative disorder chronic with eosinophilia. Translocation t(5;15)(q33;q22) with PDGFRB creating a TP53BP1-PDGFRB fusion protein.
  • 序列相似性

    Contains 2 BRCT domains.
  • 翻译后修饰

    Asymmetrically dimethylated on Arg residues by PRMT1. Methylation is required for DNA binding.
    Phosphorylated at basal level in the absence of DNA damage. Hyper-phosphorylated in an ATM-dependent manner in response to DNA damage induced by ionizing radiation. Hyper-phosphorylated in an ATR-dependent manner in response to DNA damage induced by UV irradiation.
  • 细胞定位

    Nucleus. Chromosome > centromere > kinetochore. Associated with kinetochores. Both nuclear and cytoplasmic in some cells. Recruited to sites of DNA damage, such as double stand breaks. Methylation of histone H4 at 'Lys-20' is required for efficient localization to double strand breaks.
  • Target information above from: UniProt accession Q12888 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 7158 Human
    • Entrez Gene: 27223 Mouse
    • Omim: 605230 Human
    • SwissProt: Q12888 Human
    • SwissProt: P70399 Mouse
    • Unigene: 440968 Human
    • Unigene: 383499 Mouse
    • Unigene: 481841 Mouse
    • 别名

      • 53 BP1 antibody
      • 53BP1 antibody
      • FLJ41424 antibody
      • MGC138366 antibody
      • p202 antibody
      • p53 binding protein 1 antibody
      • p53 BP1 antibody
      • p53-binding protein 1 antibody
      • p53BP1 antibody
      • TP53 BP1 antibody
      • TP53B_HUMAN antibody
      • Tp53bp1 antibody
      • TRP53 BP1 antibody
      • Tumor protein 53 binding protein 1 antibody
      • Tumor protein p53 binding protein 1 antibody
      • Tumor suppressor p53 binding protein 1 antibody
      • Tumor suppressor p53-binding protein 1 antibody
      see all

    图片

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)

      Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling 53BP1 with ab21083 at 1/4000 dilution. Nuclear staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

    • Western blot - Anti-53BP1 antibody (ab21083)
      Western blot - Anti-53BP1 antibody (ab21083)
      All lanes : Anti-53BP1 antibody (ab21083) at 1/500 dilution

      Lane 1 : 53BP1 shRNA non-transfected HeLa whole cell extracts
      Lane 2 : 53BP1 shRNA transfected HeLa whole cell extracts

      Lysates/proteins at 50 µg per lane.

      Secondary
      All lanes : HRP-conjugated anti-rabbit IgG antibody

      Predicted band size: 220 kDa
      Observed band size: 350 kDa why is the actual band size different from the predicted?



      5% SDS-PAGE

    • Western blot - Anti-53BP1 antibody (ab21083)
      Western blot - Anti-53BP1 antibody (ab21083)
      All lanes : Anti-53BP1 antibody (ab21083) at 1/2000 dilution

      Lane 1 : 293T whole cell lysate/extract
      Lane 2 : A431 whole cell lysate/extract
      Lane 3 : HeLa whole cell lysate/extract
      Lane 4 : HepG2 whole cell lysate/extract
      Lane 5 : A375 whole cell lysate/extract

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : HRP-conjugated anti-rabbit IgG antibody

      Predicted band size: 220 kDa
      Observed band size: 350 kDa why is the actual band size different from the predicted?



      5% SDS-PAGE.

      Running conditions: 80V, 15min; 140V, 40min.

      Transfer condition: Semi-dry, 18 V, 60min (Nitrocellulose membrane).

      Blocking condition: 5% non-fat milk in TBST, RT, 60min.

      Primary antibody incubation: 1/2000, 4?, overnight.

      Secondary antibody incubation: Rabbit IgG antibody (HRP), 1/10,000, RT, 1hr. 

      Washing condition: 5 ml TBST, 4 x 5min.

      Exposure system: Trident plus Western HRP Substrate.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)

      Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling 53BP1 with ab21083 at 1/4000 dilution. Nuclear staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

    • Western blot - Anti-53BP1 antibody (ab21083)
      Western blot - Anti-53BP1 antibody (ab21083)
      All lanes : Anti-53BP1 antibody (ab21083) at 1/500 dilution

      Lane 1 : Neuro2A whole cell extracts
      Lane 2 : C8D30 whole cell extracts
      Lane 3 : NIH-3T3 whole cell extracts

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : HRP-conjugated anti-rabbit IgG antibody

      Predicted band size: 220 kDa
      Observed band size: 350 kDa why is the actual band size different from the predicted?



      5% SDS-PAGE

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (ab21083)
      ab21083 (2µg/ml) staining 53BP1 in human Brain: Cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is ubiquitous nuclear staining throughout.
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

    实验方案

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (104)

    发表研究结果有使用 ab21083?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab21083 被引用在 104 文献中.

    • Bhin J  et al. Multi-omics analysis reveals distinct non-reversion mechanisms of PARPi resistance in BRCA1- versus BRCA2-deficient mammary tumors. Cell Rep 42:112538 (2023). PubMed: 37209095
    • Wang Y  et al. Wnt3a signaling with serum supply induces replication stress in cultured cells. Biochem Biophys Rep 35:101499 (2023). PubMed: 37601449
    • Tirado-Class N  et al. PHIP variants associated with Chung-Jansen syndrome disrupt replication fork stability and genome integrity. Cold Spring Harb Mol Case Stud 8:N/A (2022). PubMed: 35863899
    • Feng YL  et al. DNA nicks induce mutational signatures associated with BRCA1 deficiency. Nat Commun 13:4285 (2022). PubMed: 35879372
    • Hu X  et al. KMT2C-deficient tumors have elevated APOBEC mutagenesis and genomic instability in multiple cancers. NAR Cancer 4:zcac023 (2022). PubMed: 35898555
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 11 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-53BP1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (A549 (NSCLC))
    Permeabilization
    No
    Specification
    A549 (NSCLC)
    Blocking step
    10% FCS + 0.2% Triton X-100 as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Fixative
    Methanol
    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.

    Simon Deycmar

    Verified customer

    提交于 Feb 21 2019

    Western blot abreview for Anti-53BP1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (head and neck cancer)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (8)
    Loading amount
    15 µg
    Treatment
    drug 24h
    Specification
    head and neck cancer
    Blocking step
    BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°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 Oct 06 2015

    Western blot abreview for Anti-53BP1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MRC5VA)
    Loading amount
    15 µg
    Specification
    MRC5VA
    Gel Running Conditions
    Reduced Denaturing (8%)
    Blocking step
    Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°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 19 2011

    Immunocytochemistry/ Immunofluorescence abreview for Anti-53BP1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (T lymphocyte)
    Specification
    T lymphocyte
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% triton
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · 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 Nov 29 2010

    Question

    Dear,

    Thank you very much for your email. Earlier this year (when the antibody was still working), I was using 0.5% TritonX only for permeabilizing and 0.1% Tween for washes like you pointed out. Since this did not work recently, I was troubleshooting and switched to using stronger detergent for washes (since we were wondering whether permeabilization was the problem). All the detergents were diluted in PBS.

    We have many antibodies in the same freezer and we use them routinely for stainings. The reason is used another antibody (yH2AX) in this assay was to rule out any problem with the freezer among other possible technical issues (please see the image I had sent in the previous email). But since there is no change in the quality of stainings with yH2AX (we have had this antibody for over 3 years now), I really suspect that we might have received a faulty vial of 53BP1. I would be grateful if you could send us a new vial, the address you have mentioned is correct.

    Thank you once again for your help.

    Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 29 2012

    Answer

    Thank you very much for your reply.

    That is no problem to arrange. I have organised a new order for you for a new vial of ab21083. This is on the order number xxxx(Purchase order numberxxxxx). Please let me know if you have any problems receiving this.

    I hope this one performs better for you. However, if you do continue to have problems please do let me know.

    Until then, I wish you all the best with your research.

    Read More

    Abcam Scientific Support

    回复于 Oct 29 2012

    Question

    Dear xxxx,

    Thanks for your email. I’ve copied the questionnaire below and also attached the images for your reference.1) Abcam product code ab 21083

    2) Abcam order reference number 1090459

    3) Description of the problem Antibody does not work any more

    4) Sample preparation:
    Species
    Type of sample: cells in culture, U2OS
    Sample preparation standard immunofluorescence
    Positive control Irradiated cells (10 Gy IR)
    Negative control Untreated cells

    5) Fixation step
    Yes
    If yes: Fixative agent and concentration 3.7% Formaldehyde
    Fixation time 10 minutes
    Fixation temperature Room temperature

    6) Antigen retrieval method

    7) Permeabilization method:
    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
    Permeabilizing agent and concentration: 0.5% TritonX 100


    8) Blocking agent (eg BSA, serum…):
    Concentration 1% BSA
    Blocking time 30 minutes
    Blocking temperature Room temperature

    9) Endogenous peroxidases blocked?
    Endogenous biotins blocked?

    10) Primary antibody (If more than one was used, describe in “additional notes”) : 53BP1 (abcam) and yH2AX (millipore)
    Concentration or dilution 53BP1 1:500, yH2AX 1:250
    Diluent buffer 0.5% TritonX 100
    Incubation time 2.5 hours room temperature or Overnight at 4 degrees (tried both conditions)

    11) Secondary antibody:
    Species: Donkey
    Reacts against: Rabbit
    Concentration or dilution 1:500
    Diluent buffer 0.5% TritonX 100
    Incubation time 1 hour room temperature
    Fluorochrome or enzyme conjugate Alexa Fluor 594

    12) Washing after primary and secondary antibodies:
    Buffer 0.5% TritonX100
    Number of washes 3

    13) Detection method Immunofluorescent microscope (Leica)

    14) How many times have you run this staining? 4
    Do you obtain the same results every time? No, obtained same results in June and now it stopped working
    What steps have you altered to try and optimize the use of this antibody? Increasing concentration of Ab, increasing incubation times, permeabilizing with alternative reagents, increasing exposure time on microscope, increasing dosage of IR


    Thanks for your help!

    Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 29 2012

    Answer

    Thank you for taking time to complete our questionnaire. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. I cannot explain why the activity of the antibody diminished over the time you had it.

    One thing I would check is that you do not have a self-defrosting freezer. These automatically defrost, raising and lowering the temperature, forcing the antibody through freeze-thaw cycles, potentially decreasing its activity.

    Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. There is only one thing I would point out which may be worth taking into consideration. You have used 0.5% TritonX in the antibody diluent as well as the wash buffers. This as well as the permiabilisation step may be too strong. I would use one 10 minute permiabilisation step (using 0.2% Triton X in PBS) at room temperature. Following this I would only use 0.1% Tween 20 in your diluent buffers. You did not mention explicitly but are you using PBS or TBS for the diluent buffers?

    If you would like, I can send you a new vial of ab21083 to be sent to you to see if this makes a difference in your staining? If you would like for me to arrange this please could you confirm I have the correct addressbr/>Alternatively, if you would prefer I can offer a credit note, or refund in compensation.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Abcam Scientific Support

    回复于 Oct 29 2012

    Question

    Phone call reporting change in activity of the antibody following freezing.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 25 2012

    Answer

    Thank you for contacting us yesterday and sorry for the delay in getting back to you.

    I am sorry to hear that you have been having some difficulty in using the Anti-53BP1 antibody (ab21083). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    As we discussed on the phone, I am attaching our questionnaire so that I can gather further information regarding the samples tested and the protocol used. Once I have receive the completed questionnaire, I will look at the protocol and see if there are any suggestions I can make that may improve the results.

    If you could include some images of the staining when the antibody worked and how the latest experiments have been that would be very helpful.

    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

    Abcam Scientific Support

    回复于 Oct 25 2012

    Question

    Phone call requesting the immunogen sequence used to raise anti-53BP1 antibody (ab21083).

    Read More

    Abcam community

    Verified customer

    Asked on May 16 2012

    Answer

    Thank you for your inquiry.

    Unfortunately, we are not able to release the immunogen sequence for this particular antibody as it is proprietary information. However, I can tell you that it falls within the carboxy-terminus of h53BP1.

    We aim to provide as much information as possible to our customers, so I am sorry that this has not been possible on this occasion.

    Should you have any further questions, please do not hesitate to contact us.

    Read More

    Abcam Scientific Support

    回复于 May 16 2012

    Question

    Dear Technical, please find attached antibody data for ab21083 , and advise what your suggestion maybe.

    Thanks

    Cat.#: ab21083
    Lot. # GR56537-11
    1. Order details:
    - Antibody storage conditions (temperature/reconstitution etc)
    -20oC
    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: human cancer cells
    - Cell extract or Nuclear extract: Nuclear extract
    - Purified protein or Recombinant protein: Purified protein
    4. The lysate
    - How much protein was loaded: 20ul
    - What lysis buffer was used: NP-40 lysis buffer
    - What protease inhibitors were used: cocktail PMSF
    - What loading buffer was used: SDS-PAGE loading buffer 5X
    - Did you heat the samples: temperature and time: yes ,95o, 8min
    5. Electrophoresis/Gel conditions/ Transfer conditions
    - Reducing or non reducing gel: non reducing
    - Gel percentage : 10% or 15%
    - Transfer conditions: wet transfer 300mA 90min

    6. Blocking conditions
    - Buffer: TBST
    - Blocking agent: milk, BSA, serum, what percentage: no-fat milk 5%
    - Incubation time:90min
    - Incubation temperature RT:
    7. Primary Antibody
    - Specification (in which species was it raised against): Hu Ms
    - At what dilution(s) have you tested this antibody:
    - What dilution buffer was used: TBST
    - Incubation time: over night
    - Incubation temperature: 4oC
    - What washing steps were done: TBST 10minx3
    8. Secondary Antibody
    - Specification (in which species was it raised against)? rabbit
    - At what dilution(s) have you tested this antibody:
    - Incubation time:2h
    - Wash steps: TBST 10minx3
    - Do you know whether the problems you are experiencing come from the secondary?
    9. Detection method
    ECl, ECl+, other detection method: ECl+ santa
    10. Background bands
    - Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control):
    yes, we have run a “No primary” control
    - Is the blocking step sufficient? Yes , 2h RT
    - Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps)
    - At what size are the bands migrating? Could they be degradation products of your target?
    - Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient)
    11. Did you apply positive and negative controls along with the samples? Please specify.
    12. Optimization attempts
    - How many times have you tried the Western? 3
    - Do you obtain the same results every time e.g. are background bands always in the same place?
    No bands
    - What steps have you altered?

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    Abcam community

    Verified customer

    Asked on Mar 12 2012

    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hearthe customer has haddifficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    As far as I know, Biomoles have been made aware that they are not an authorized distributor of Abcam products. Regrettably, I am sorry this means we are not able to provide replacements or credit notes for faulty items purchased and resold by Biomoles. However, we are pleased to help as much as we can with technical support. Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

    1. I recommend to check the amount of protein loaded. Try a protein assay on the sample and load 20 -30 ug of total protein per lane of the gel to ensure there is enough protein for detection.

    2. Please confirm what type of cancer cells are being tested? What positive controls have been used? For example, HeLa whole cell lysate would be a suitable positive control.

    3. Could you confirm if the transfer of protein to the membrane and quality of the sample has been assessed using a loading control?

    4. Please confirm what dilutions of antibody have been tried? This may require some optimization.

    5. Is the secondary antibody working well with other antibodies?

    I hope this information is helpful. Thank you for your cooperation and for your understanding of our policy regarding unofficial distributors. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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    Abcam Scientific Support

    回复于 Mar 12 2012

    Question

    Why is the molecular weight band higher than expected?

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    Abcam community

    Verified customer

    Asked on Nov 17 2008

    Answer

    Thank you for your enquiry. We do notice that there is some size discrepancy between the calculated MW vs. observed MW with the ab21083 antibody. This product has been tested in house on Hela cell lysate and the antigen that was used to generate the antibody by ELISA and WB. We also found that the peptide blocks antibody detection by WB. Based on our testing and data collected we concluded and feel confident that the antibody is detecting the 53BP1 protein. I have included PubMed IDs for several publications which describe detection of the 53BP1 protein by WB. The literatures indicate that 53BP1 protein migrates significantly slower than its predicted size. PubMed ID: 11238909 11134068 9748285 15840649 This product is one of our top selling antibodies and we have not received complaints regarding detection of the incorrect protein. This is currently all the information we have available. But if we do happen to obtain more data, we will be sure to forward you the information.

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    Abcam Scientific Support

    回复于 Nov 17 2008

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