概述

  • 产品名称Anti-PRPF8抗体
    参阅全部 PRPF8 一抗
  • 描述
    兔多克隆抗体to PRPF8
  • 经测试应用适用于: IHC-P, WB, ICC/IF, IPmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Cow
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human PRPF8.

    (Peptide available as ab86623.)

  • 阳性对照
    • This antibody gave a positive signal in the following cell lysates: HeLa (Whole Cell lysate and Nuclear extract); NIH 3T3; A431; MEF1.

性能

应用

Our Abpromise guarantee covers the use of ab79237 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
IHC-P Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 250 kDa (predicted molecular weight: 274 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml.

靶标

  • 功能Central component of the spliceosome, which may play a role in aligning the pre-mRNA 5'- and 3'-exons for ligation. Interacts with U5 snRNA, and with pre-mRNA 5'-splice sites in B spliceosomes and 3'-splice sites in C spliceosomes.
  • 组织特异性Widely expressed.
  • 疾病相关Defects in PRPF8 are the cause of retinitis pigmentosa type 13 (RP13) [MIM:600059]. RP leads to degeneration of retinal photoreceptor cells. Patients typically have night vision blindness and loss of midperipheral visual field. As their condition progresses, they lose their far peripheral visual field and eventually central vision as well. RP13 inheritance is autosomal dominant.
  • 序列相似性Contains 1 MPN (JAB/Mov34) domain.
  • 结构域The MPN domain has structural similarity with viral ribonucleases and RNase H, but unlike RNases, it does not bind any metal ions.
  • 翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 细胞定位Nucleus speckle.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 220 kDa U5 snRNP specific protein antibody
    • 220 kDa U5 snRNP-specific protein antibody
    • Apoptosis regulated protein 1 antibody
    • Apoptosis regulated protein 2 antibody
    • HPRP8 antibody
    • p220 antibody
    • Pre mRNA processing factor 8 antibody
    • Pre mRNA-processing factor 8, S. cerevisiae, homolog of antibody
    • Pre-mRNA-processing-splicing factor 8 antibody
    • Precursor mRNA processing protein antibody
    • PRP8 antibody
    • PRP8 homolog antibody
    • PRP8 pre mRNA processing factor 8 homolog antibody
    • PRP8_HUMAN antibody
    • PRPC8 antibody
    • Prpf8 antibody
    • Retinitis pigmentosa 13 (autosomal dominant) antibody
    • RP13 antibody
    • SNRNP220 antibody
    • Splicing factor Prp8 antibody
    • U5 snRNP specific protein antibody
    • U5 snRNP specific protein (220 kD), ortholog of S. cerevisiae Prp8p antibody
    see all

Anti-PRPF8 antibody 图像

  • All lanes : Anti-PRPF8 antibody (ab79237) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 274 kDa
    Observed band size : 250 kDa (why is the actual band size different from the predicted?)
    The 250 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to PRPF8.
  • IHC image of PRPF8 staining in human colon carcinoma 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 ab79237, 10µg/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.
  • ICC/IF image of ab79237 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79237, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.
  • PRPF8 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to PRPF8 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab79237.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 250kDa; PRPF8

Anti-PRPF8 antibody (ab79237)参考文献

This product has been referenced in:
  • van Maldegem F  et al. CTNNBL1 facilitates the association of CWC15 with CDC5L and is required to maintain the abundance of the Prp19 spliceosomal complex. Nucleic Acids Res 43:7058-69 (2015). WB ; Mouse . Read more (PubMed: 26130721) »
  • Laetsch TW  et al. Multiple components of the spliceosome regulate Mcl1 activity in neuroblastoma. Cell Death Dis 5:e1072 (2014). WB . Read more (PubMed: 24556687) »

See all 2 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Mouse Cell lysate - whole cell (C2C12)
Specification C2C12
Treatment control or PRPF8 siRNA
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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提交于 Oct 14 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Sample Human Cell (HaCaT)
Specification HaCaT
Permeabilization Yes - Triton X-100 plus
Fixative Paraformaldehyde
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提交于 Oct 11 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (HEK293 cells)
Specification HEK293 cells
Treatment control or PRPF8 siRNA treated (300pM per 10 cm plate, 48h)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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提交于 Oct 11 2013

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring...

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I suggest trying anti-ATE1 ab90561, which is a mouse monoclonal, tested for reactivity with human Jurkat but untested on mouse samples. One issue might be reactivity of an anit-mouse IgG secondary with endogenous IgG in the mosue lysate, but this is un...

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We have two other antibodies against each of these targets which may give you a better result. However, before sending replacements, we typically collect details of the protocols from our customers before issuing a replacement or credit/refund, in t...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Xenopus laevis Cell (egg extract with sperm nuclei)
Specification egg extract with sperm nuclei
Fixative Formaldehyde
Permeabilization Yes - methanol
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
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提交于 Oct 27 2011

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Cell lysate - whole cell (egg extract)
Loading amount 40 µg
Specification egg extract
Gel Running Conditions Non-reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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提交于 Oct 17 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"