Anti-NFYB/CBF-B抗体(ab6559)

Submit a review Q&A (2)References (4)

Key features and details

  • Rabbit polyclonal to NFYB/CBF-B
  • Suitable for: EMSA
  • Reacts with: Mouse, Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-NFYB/CBF-B抗体
    参阅全部 NFYB/CBF-B 一抗

  • 描述

    兔多克隆抗体to NFYB/CBF-B

  • 宿主

    Rabbit

  • 经测试应用

    适用于: EMSAmore details

  • 种属反应性

    与反应: Mouse, Human

  • 免疫原

    Synthetic peptide corresponding to NFYB/CBF-B (N terminal) conjugated to keyhole limpet haemocyanin.
    Sequence:

    QQISGVQQIQFS


    Database link: NP_000729.2

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

  • 常规说明

     This product was previously labelled as NFYB

     

性能

应用

Our Abpromise guarantee covers the use of ab6559 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
EMSA Use 2µl for 104 cells.

靶标

  • 功能

    Stimulates the transcription of various genes by recognizing and binding to a CCAAT motif in promoters, for example in type 1 collagen, albumin and beta-actin genes.

  • 序列相似性

    Belongs to the NFYB/HAP3 subunit family.

  • 结构域

    Can be divided into 3 domains: the weakly conserved A domain, the highly conserved B domain thought to be involved in subunit interaction and DNA binding, and the Glu-rich C domain.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • CAAT box DNA binding protein subunit B antibody
      • CAAT box DNA-binding protein subunit B antibody
      • CBF A antibody
      • CBF B antibody
      • CCAAT binding transcription factor subunit A antibody
      • HAP 3 antibody
      • HAP3 antibody
      • NF Y protein chain B antibody
      • NF YB antibody
      • NF-YB antibody
      • NFYB antibody
      • NFYB_HUMAN antibody
      • Nuclear transcription factor Y beta antibody
      • Nuclear transcription factor Y subunit B antibody
      • Nuclear transcription factor Y subunit beta antibody
      • Transcription factor NF Y B subunit antibody
      see all

    图片

    • Electrophoretic Mobility Shift Assay - Anti-NFYB/CBF-B antibody (ab6559)
      Electrophoretic Mobility Shift Assay - Anti-NFYB/CBF-B antibody (ab6559)Image from Grskovic M et al., PLoS Genet. 2007 Aug;3(8):e145. Fig 6.; doi:10.1371/journal.pgen.0030145; August 31, 2007, PLoS Genet 3(8): e145.

      EMSA was performed with a double-stranded, biotin-labeled oligonucleotide in the presence of murine ES cell nuclear extracts and increasing amounts of anti-NFYB/CBF-B antibody (ab6559; 2 µg or 6 µg) or unrelated antibody.

    实验方案

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    文献 (4)

    发表研究结果有使用 ab6559？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab6559 被引用在 4 文献中.

    • Braden CR & Neufeld TP Atg1-independent induction of autophagy by the Drosophila Ulk3 homolog, ADUK. FEBS J 283:3889-3897 (2016). PubMed: 27717182
    • Su B  et al. Interleukin-1ß/Iinterleukin-1 receptor-associated kinase 1 inflammatory signaling contributes to persistent Gankyrin activation during hepatocarcinogenesis. Hepatology 61:585-97 (2015). PubMed: 25294684
    • Huang Y  et al. Phospho-?Np63a is a key regulator of the cisplatin-induced microRNAome in cancer cells. Cell Death Differ 18:1220-30 (2011). WB ; Human . PubMed: 21274007
    • Grskovic M  et al. Systematic identification of cis-regulatory sequences active in mouse and human embryonic stem cells. PLoS Genet 3:e145 (2007). EMSA ; Mouse . PubMed: 17784790

    客户评价及客户问答

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    Question

    1. Antibody storage conditions (temperature/reconstitution etc) -20oC 2. Please describe the problem (high background, wrong band size, more bands, no band etc). major band is wrong size 50kd 3. On what material are you testing the antibody in WB? Species: 293T human Cell extract or Nuclear extract:Cell extract Purified protein or Recombinant protein: 3. The lysate How much protein was loaded:100ug What lysis buffer was used:RIPA What protease inhibitors were used:no What loading buffer was used: 3X loading dye from moleunlar cloning Did you heat the samples: temperature and time:95 oC , 10min 4. Electrophoresis/Gel conditions/ Transfer conditions Reducing or non reducing gel: reducing gel Gel percentage :15% Transfer conditions: 150V 2hr Buffer:5% milk Blocking agent: milk, BSA, serum, what percentage:5% Incubation time:over night Incubation temperature:4 oC 6. Primary Antibody Specification (in which species was it raised against):rabbit At what dilution(s) have you tested this antibody:1:1000 What dilution buffer was used:5% milk Incubation time:3hr Incubation temperature:RT What washing steps were done:1X TBST 3 times 7. Secondary Antibody Specification (in which species was it raised against)?rabbit At what dilution(s) have you tested this antibody:1:2000 Incubation time1hr Wash steps:1X TBST 3times Do you know whether the problems you are experiencing come from the secondary?the secondary ab is ok 8. 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): Is the blocking step sufficient? 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. 10. Optimization attempts How many times have you tried the Western?Twice Do you obtain the same results every time e.g. are background bands always in the same place? yes What steps have you altered? 1o ab incubation time

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    Answer

    I'm sorry to hear you are having problems with ab6559. The protein NF-YB is know to function as a heterotrimeric transcription factor when bound to NF-YA and NF-YC. It is therefore possible that the large protein observed is made of NF-YB and other parts of the heterotrimer (NF-YB is 22.8kDa, NF-YA is 36kDa, NF-YC is 50.3kDa). I would recommend the following modifications to your protocol: -use protease inhibitors in the lysis buffer -make sure the loading buffer is reducing (i.e. containing SDS, beta mercaptoethanol, DTT) -load less protein per well (25-40ug) -block the membrane in milk (you may wish to try BSA 5% as well) for 1hr at RT -incubate the primary antibody 1:1000 at 4C overnight in TBST (no milk) -incubate the secondary antibody in TBST Please note that the protein is found in the nucleus and should the band be due to non-specific binding of the antibody you may need to concentrate your samples by nuclear extraction prior to SDS separation. I hope these suggestions will help you, please do not hesitate to contact us again if you still experience problems,

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    Question

    I recently purchased NF-YB for testing in Arabidopsis. The reference you provide (EMBO J. 13:4042-4053) does not identify the peptide used for the detection of NF-YB, but instead references another paper: Mantovani et al (1992) EMBO J. 11:3315-3322. This paper says somewhat unclearly, that a peptide from position 200-897 was used to raise their polyclonal antibodies and they supply a third paper as a reference for those amino acids. On your datasheet you mention a peptide corresponding to a region near the N-terminus of the human protein. Position 200-897 doesn't really sound like the "N-terminus", so I find myself confused. Could you please tell me the actual peptide used to raise the NF-YB antibody?

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    Answer

    The peptide sequence is: QQI SGV QQI QFS

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