PRPF8 Recombinant Rabbit Monoclonal Antibody [JG57-38]
Catalog# ET7108-18
PRPF8 Recombinant Rabbit Monoclonal Antibody [JG57-38]
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WB
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IF-Cell
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
PRPF8 Recombinant Rabbit Monoclonal Antibody [JG57-38]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human PRPF8 aa 2,200-2,335 / 2,335.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC
Molecular Weight
Predicted band size: 274 kDa
Positive Control
SiHa cell lysate, A549 cell lysate, 293T, LOVO, SiHa, rat brain tissue, human colon tissue, human prostate carcinoma tissue, mouse testis tissue, A549.
Conjugation
unconjugated
Clone Number
JG57-38
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IF-Cell
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1:50-1:200
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
Target
Function
PRP8, also designated pre-mRNA-processing-splicing factor 8, is a highly conserved nuclear protein and a central component of the catalytic core of the spliceosome, where it may be involved in various molecular rearrangements. PRP8, which is widely expressed, plays a role in transesterification reactions that regulate splicesome-induced pre-mRNA splicing. Specifically, PRP8 interacts with the GU dinucleotide at the 5' splice site (5'SS) and forms a specific UV-inducible cross-link. It also interacts functionally with the 3'SS, affecting the efficiency of the second catalytic step. PRP8 may play a role in the first transesterification step, as PRP8 mutations that prohibit negative regulation of PRP28 or PRP44/Brr2 subsequently block U4 activation. In addition, PRP8 interacts with a conserved region of U6 that is instrumental in the formation of the catalytic core of the spliceosome.
Background References
1. Song E J et al. The Prp19 complex and the Usp4Sart3 deubiquitinating enzyme control reversible ubiquitination at the spliceosome. Genes Dev 24:1434-1447 (2010).
2. McKie A B et al. Mutations in the pre-mRNA splicing factor gene PRPC8 in autosomal dominant retinitis pigmentosa (RP13). Hum Mol Genet 10:1555-1562 (2001).
Tissue Specificity
Widely expressed.
Subcellular Location
Nucleus. Spliceosome.
Synonyms
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
ExpandImages
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ICC staining of PRPF8 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of PRPF8 in LOVO cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of PRPF8 in SiHa cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-PRPF8 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-PRPF8 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-PRPF8 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-PRPF8 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of PRPF8 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7108-18, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).
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Western blot analysis of PRPF8 on different lysates with Rabbit anti-PRPF8 antibody (ET7108-18) at 1/2,000 dilution.
Lane 1: 293T (Human embryonic kidney cell) cell lysate
Lane 2: HepG2 (Human hepatocellular carcinoma cell) cell lysate
Lane 3: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 4: RAW264.7 (Mouse monocytic macrophage leukemia cell) cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 180 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: ET7108-18, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 274 kDa
Observed band size: 274 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"