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Anti-SC35抗体(ab28428)

Reviews (2)Q&A (7)References (9)
Western blot - Anti-SC35 antibody (ab28428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SC35 antibody (ab28428)

Key features and details

  • Rabbit polyclonal to SC35
  • Suitable for: WB, IHC-P
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-SC35 antibody [EPR12238] (ab204916)
  • 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-SC35抗体
    参阅全部 SC35 一抗

  • 描述

    兔多克隆抗体to SC35

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IHC-Pmore details

  • 种属反应性

    与反应: Human, Recombinant fragment

  • 免疫原

    Recombinant full length protein (Human)

  • 阳性对照

    • HeLa nuclear extract

  • 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

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab28428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
1/2000 - 1/5000. Detects a band of approximately 30 kDa (predicted molecular weight: 26 kDa).
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
说明
WB
1/2000 - 1/5000. Detects a band of approximately 30 kDa (predicted molecular weight: 26 kDa).
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

靶标

  • 功能

    Necessary for the splicing of pre-mRNA. It is required for formation of the earliest ATP-dependent splicing complex and interacts with spliceosomal components bound to both the 5'- and 3'-splice sites during spliceosome assembly. It also is required for ATP-dependent interactions of both U1 and U2 snRNPs with pre-mRNA. Interacts with other spliceosomal components, via the RS domains, to form a bridge between the 5'- and 3'-splice site binding components, U1 snRNP and U2AF. Binds to purine-rich RNA sequences, either 5'-AGSAGAGTA-3' (S=C or G) or 5'-GTTCGAGTA-3'. Can bind to beta-globin mRNA and commit it to the splicing pathway.

  • 序列相似性

    Belongs to the splicing factor SR family.
    Contains 1 RRM (RNA recognition motif) domain.

  • 翻译后修饰

    Extensively phosphorylated on serine residues in the RS domain.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • 35 kDa antibody
      • arginine/serine-rich 2 antibody
      • PR264 antibody
      • Protein PR264 antibody
      • SC 35 antibody
      • SC-35 antibody
      • SC35 antibody
      • Serine/arginine-rich splicing factor 2 antibody
      • SFRS 2 antibody
      • SFRS2 antibody
      • SFRS2A antibody
      • Splicing component 35 kDa antibody
      • Splicing component antibody
      • Splicing factor antibody
      • Splicing factor arginine/serine rich 2 antibody
      • Splicing factor SC35 antibody
      • Splicing speckle antibody
      • Splicing speckles antibody
      • SR splicing factor 2 antibody
      • SRp30b antibody
      • SRSF2 antibody
      • SRSF2_HUMAN antibody
      see all

    图片

    • Western blot - Anti-SC35 antibody (ab28428)
      Western blot - Anti-SC35 antibody (ab28428)
      All lanes : Anti-SC35 antibody (ab28428)

      Lane 1 : recombinant SC35 protein
      Lane 2 : HeLa nuclear lysate

      Predicted band size: 26 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SC35 antibody (ab28428)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SC35 antibody (ab28428)
      IHC image of ab28428 staining in human seminoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28428, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    文献 (9)

    发表研究结果有使用 ab28428？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab28428 被引用在 9 文献中.

    • Yadav P  et al. Hypoxia-induced loss of SRSF2-dependent DNA methylation promotes CTCF-mediated alternative splicing of VEGFA in breast cancer. iScience 26:106804 (2023). PubMed: 37235058
    • Zhu Z  et al. Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis. Onco Targets Ther 13:11301-11313 (2020). PubMed: 33177838
    • Ilik IA  et al. SON and SRRM2 are essential for nuclear speckle formation. Elife 9:N/A (2020). PubMed: 33095160
    • Komeno Y  et al. SRSF2 Is Essential for Hematopoiesis, and Its Myelodysplastic Syndrome-Related Mutations Dysregulate Alternative Pre-mRNA Splicing. Mol Cell Biol 35:3071-82 (2015). PubMed: 26124281
    • Erkelenz S  et al. Balanced splicing at the Tat-specific HIV-1 3'ss A3 is critical for HIV-1 replication. Retrovirology 12:29 (2015). PubMed: 25889056
    • Rampal R  et al. DNA hydroxymethylation profiling reveals that WT1 mutations result in loss of TET2 function in acute myeloid leukemia. Cell Rep 9:1841-55 (2014). PubMed: 25482556
    • Berulava T  et al. FTO levels affect RNA modification and the transcriptome. Eur J Hum Genet : (2012). PubMed: 22872099
    • Kralovicova J  et al. Compensatory signals associated with the activation of human GC 5' splice sites. Nucleic Acids Res : (2011). PubMed: 21609956
    • Toyota M  et al. Epigenetic silencing of microRNA-34b/c and B-cell translocation gene 4 is associated with CpG island methylation in colorectal cancer. Cancer Res 68:4123-32 (2008). WB ; Human . PubMed: 18519671

    客户评价及客户问答

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    1-9 of 9 Abreviews or Q&A

    Western blot abreview for Anti-SC35 antibody

     Inconclusive
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Bone marrow cell)
    Gel Running Conditions
    Reduced Denaturing (Any kDMini-PROTEAN TGX Gel)
    Loading amount
    500000 cells
    Specification
    Bone marrow cell
    Blocking step
    Milk 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 Jun 10 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SC35 antibody

     Inconclusive
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (A549 cells (alveolar basal epithelial cells))
    Specification
    A549 cells (alveolar basal epithelial cells)
    Fixative
    Formaldehyde
    Permeabilization
    Yes - 0.5% Triton X-100
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · 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 Jan 24 2011

    Question

    Ab28428 gave no bands in Western blot.

    Read More
    Answer

    Thank you for contacting us.

    Your credit note ID is ***


    I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone at 888-772-2226. Please refer to the credit note ID in any correspondence with our accounting department.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

    Question

    If we could get the replacement products, that would fantastic. Is shipping on ice standard, or are there any cases that the antibody would ship on dry ice or a similar cooling agent? If the anti-ZRSR2 antibody is shipped on Tuesday (the 28th is Memorial Day), when would it be expected to arrive here in Seattle?

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry these products did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued free of charge replacements with the order number XXXX. These products are expected to arrive to us early next week and we will ship them to you by overnight delivery. These product will the shipped on blue ice as this condition is suitable for the majority of our products.To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Question

    I think there was some confusion somewhere, sorry about that. Under all conditions that we have tried in our lab we have not seen bands corresponding to the protein on our Western blots, for both anti-SC35 and anti-ZRSR2. We have separately used other antibodies on the same blots that the anti-SC35 and anti-ZRSR2 did not work on and seen product bands for those proteins (not the two on this inquiry). We have not had luck with the ab28428 and ab50753 in our hands. Again I apologize for any confusion.

    Read More
    Answer

    Thank you for clarifying this for me.

    As you have used these products in applications and species listed on the datasheet and as you have attempted extensive troubleshooting and I can see no further advice which may help in this instance, we believe that the issue may indeed lie in the vials which you have received. It may or may not be shipping or handling related as both products were from the same shipment. Regardless, these products are covered by our Abpromise guarantee. Under this guarantee I may replace or refund these products for you.

    While we currently have a new lot of our anti-SC35 antibody (ab28428) available. There is a slight delay with our anti-ZRSR2 antibody as we are awaiting a new lot of this product. The expected arrival of this product is 28 May.

    If you would like, I can arrange these replacements to be sent to you free of charge. Please know that these products would also be covered by our Abpromise should you experience any further issues we will replace them or refund the original purchase.

    Please let me know how you would like to proceed. Also do not hesitate to contact me with any questions you may have.

    Read More

    Question

    Even with using the higher concentrations as previously described, there are no bands even after the longest development times. I should note that one of the two anti-bodies (alpha-tubulin) that did work on the blot was more than 10 fold more dilute than the lowest suggested concentration for that antibody.

    Read More
    Answer

    Thank you for your response.

    I do wish to apologize for repeatedly inquiring this further information but I wanted to be clear about the issue you have been having. Based on what you have told me you have previously used the anti-SC35 antibody successfully, in gels loading 25-60ug/well of total protein. During these experiments the product did work. However, you latest experiments using this product in which you have loaded 400ng and 800ng total protein/well and used the product at 1:200 have failed. You have also seen a similar issue when using anti-ZRSR2. Have I got this correct?

    Thank you for your help in clarifying this for me. I look forward to your response and helping you to achieve your desired results. Please let me know if you have any questions.

    Read More

    Question

    The protein is in fact unpurified, whole-cell lysates. We used this higher concentration in order to make sure we could get bands with the antibodies more so than to quantify the amount of each protein in the cells. We had used the Anti-SC35 before with gels that were loaded with 15-25ug and ~60ug of protein from whole cell lysate. The two anti-bodies that were used and we got bands for were anti-alpha tubulin (1:40000) and anti-SF3B1 (1:2000) for which the same blotting and washing conditions were used; the secondary for the alpha-tubulin was anti-mouse (same concentration) and the same anti-rabbit was used for the SF3B1. For the Western that ~800ng was loaded, we didn't strip the blot. Both sets of antibodies were blotted at the same time; the blot was cut according the expected sizes and incubated with each primary separately. We have stripped blots in the past, but wanted to make the same exact protein samples were being used without having to factor in any loss due to the stripping procedure.

    Read More
    Answer

    Thank you for this further information.

    I would like to repeat that I am sorry that these products are not working as expected for you. I see that you have used the anti-SC35 antibody at a much higher concentration than we have had to use in our tests and the anti-ZRSR2 at the recommended starting dilution. My only concern is that you use much less protein than we normally recommend, which is usually ˜25-50ug per well though it may be a low as 10ug. While the levels which you use might work with the purified protein or peptide we are unsure whether there would be enough SC35 or ZRSR2 within your sample for our product to detect. When you were using these products with the higher protein levels were you able to get results?

    This product is covered by our Abpromise guarantee. If we cannot remedy this issue together we will replace or refund it under our Abpromise guarantee.

    I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

    Use our products? Submit an Abreview. Earn rewards!

    https://www.abcam.com/abreviews

    Read More

    Question

    Inquiry: We have tried using the Anti-SC35 antibody (a b28428) multiple times in our lab without any success. The antibody was ordered back in April and was delivered the following week (April 25th). It was placed in our -20 freezer when we received it. Increasing amounts of protein have been used in multiple gels, the latest of which was both 400 and 800ng of total protein from 293T and HeLa cells. The proteins were denatured and loaded on to 4-20% Tris-Glycine gels, transferred overnight at 4'C using the Criterion Blotter. The blot has been blocked with 5% non-fat dry milk in TBST at one hour at room temp, as well as overnight. When loading the 800ng, primary was used at 1:200 in TBST 5% milk and incubated overnight at 4'C, washed 3x10 minutes in TBST, incubated with secondary at 25'C for one hour (Donkey Anti-Rabbit IgG HRP from Jackson ImmnuoResearch) at 1:50,000 and in 5% milk TBST, and washed 3x 10minutes in TBST. The blots were then developed with SuperSignal West Femto ECL for five minutes, and then exposed to high-sensitivity x-ray for up to 30 minutes. No bands were seen, however the same samples (and same wash, secondary conditions) had bright, strong bands for two other proteins. Is there any chance this antibody went bad? If so, would it be possible to get a new lot of it? **The same Westerns were also blotted with Anti-ZRSR2 (a b50753), ordered at the same time as the SC35 antibody, also resulting in no bands. The lot number for this antibody was not written down before aliquoting and storage at -20. On the same gel as above, this was used at the suggested 2.5 ug/mL and incubated overnight at 4'C. The same washing, secondary and detection conditions were also used.*** Any help with this would be greatly appreciated.

    Read More
    Answer

    Thank you for contacting us.

    I am sorry to hear of the issues which you have been facing when using these products. I would like to thank you for including your protocols details with this inquiry. You state that you have use up to 800ng. Are your samples purified? If not we would recommend attempting whole cell or nuclear lysates with a total protein concentration of 20-50ug/well.

    You also mention that other antibodies used on the same blot did work. Could you tell me which antibodies these where and detail the stripping procedure which you used on the blot? Were these other antibodies used before or after the SC35 and ZRSR2 antibodies?



    These products are covered by our Abpromise guarantee. If we cannot remedy this issue we will replace or refund the product/s under our Abpromise guarantee, as you are using them according to specifications listed on our datasheet.

    I look forward to your response. Please do not hesitate to contact us if you need any advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Question

    How was this antibody tested in Western blot and what size band did you observe?

    Read More
    Answer

    Thank you for your enquiry. ab28428 is a polyclonal antibody generated from baculovirus-expressed recombinant protein and purified through a protein A column. The size of the protein (SC35) detected from HeLa cell nuclear extract is about 30kDa. I hope this information helps. Please do not hesitate to contact us if you need anything further.

    Read More

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