Anti-PRPF8/Prp8 antibody (ab79237)
Key features and details
- Rabbit polyclonal to PRPF8/Prp8
- Suitable for: IHC-P, WB, ICC/IF, IP
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-PRPF8/Prp8 antibody
See all PRPF8/Prp8 primary antibodies -
Description
Rabbit polyclonal to PRPF8/Prp8 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Cow
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Immunogen
Synthetic peptide corresponding to Human PRPF8/Prp8 aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab86623) -
Positive control
- This antibody gave a positive signal in the following cell lysates: HeLa (Whole Cell lysate and Nuclear extract); NIH 3T3; A431; MEF1.
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General notes
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
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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.
Application Abreviews Notes IHC-P Use a concentration of 10 µg/ml.WB (3) Use a concentration of 1 µg/ml. Detects a band of approximately 250 kDa (predicted molecular weight: 274 kDa).ICC/IF (2) Use a concentration of 1 µg/ml.IP Use a concentration of 5 µg/ml.Notes 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.Target
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Function
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. -
Tissue specificity
Widely expressed. -
Involvement in disease
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. -
Sequence similarities
Contains 1 MPN (JAB/Mov34) domain. -
Domain
The MPN domain has structural similarity with viral ribonucleases and RNase H, but unlike RNases, it does not bind any metal ions. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus speckle. - Information by UniProt
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Database links
- Entrez Gene: 10594 Human
- Entrez Gene: 192159 Mouse
- Omim: 607300 Human
- SwissProt: Q6P2Q9 Human
- SwissProt: Q99PV0 Mouse
- Unigene: 181368 Human
- Unigene: 3757 Mouse
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Alternative names
- 220 kDa U5 snRNP specific protein antibody
- 220 kDa U5 snRNP-specific protein antibody
- Apoptosis regulated protein 1 antibody
see all
Images
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Western blot - Anti-PRPF8/Prp8 antibody (ab79237)All lanes : Anti-PRPF8/Prp8 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
All lanes : 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/Prp8.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRPF8/Prp8 antibody (ab79237)IHC image of PRPF8/Prp8 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.
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Immunocytochemistry/ Immunofluorescence - Anti-PRPF8/Prp8 antibody (ab79237)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. -
Immunoprecipitation - Anti-PRPF8/Prp8 antibody (ab79237)PRPF8/Prp8 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to PRPF8/Prp8 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/Prp8
Protocols
Datasheets and documents
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Datasheet download
References (8)
ab79237 has been referenced in 8 publications.
- Wang S et al. Pre-mRNA Processing Factor 8 Accelerates the Progression of Hepatocellular Carcinoma by Regulating the PI3K/Akt Pathway. Onco Targets Ther 13:4717-4730 (2020). PubMed: 32547101
- Alkalay E et al. The Sub-Nuclear Localization of RNA-Binding Proteins in KSHV-Infected Cells. Cells 9:N/A (2020). PubMed: 32854341
- Schilling J et al. Deregulated Splicing Is a Major Mechanism of RNA-Induced Toxicity in Huntington's Disease. J Mol Biol N/A:N/A (2019). PubMed: 30711541
- Onyango DO et al. PRPF8 is important for BRCA1-mediated homologous recombination. Oncotarget 8:93319-93337 (2017). PubMed: 29212152
- Tarallo R et al. The nuclear receptor ERß engages AGO2 in regulation of gene transcription, RNA splicing and RISC loading. Genome Biol 18:189 (2017). WB ; Human . PubMed: 29017520
- Esquerdo M et al. Expression Atlas of the Deubiquitinating Enzymes in the Adult Mouse Retina, Their Evolutionary Diversification and Phenotypic Roles. PLoS One 11:e0150364 (2016). IHC . PubMed: 26934049
- 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 . PubMed: 26130721
- Laetsch TW et al. Multiple components of the spliceosome regulate Mcl1 activity in neuroblastoma. Cell Death Dis 5:e1072 (2014). WB . PubMed: 24556687
Images
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Western blot - Anti-PRPF8/Prp8 antibody (ab79237)All lanes : Anti-PRPF8/Prp8 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
All lanes : 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/Prp8.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRPF8/Prp8 antibody (ab79237)
IHC image of PRPF8/Prp8 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.
-
Immunocytochemistry/ Immunofluorescence - Anti-PRPF8/Prp8 antibody (ab79237)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.
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Immunoprecipitation - Anti-PRPF8/Prp8 antibody (ab79237)
PRPF8/Prp8 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to PRPF8/Prp8 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/Prp8
