PRODUCT CODE: ET7108-18

PRPF8 Recombinant Rabbit Monoclonal Antibody [JG57-38] (ET7108-18)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of PRPF8 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7108-18, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: SiHa cell lysate<br />
Lane 2: A549 cell lysate
  • Western blot analysis of PRPF8 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7108-18, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: SiHa cell lysate<br />
Lane 2: A549 cell lysate
  • 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).
  • 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).
  • 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).
  • 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) for 20 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.
  • 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) for 20 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.
  • 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) for 20 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.
  • 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) for 20 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.
  • 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).
Western blot analysis of PRPF8 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7108-18, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: SiHa cell lysate
Lane 2: A549 cell lysate

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

PRPF8 Recombinant Rabbit Monoclonal Antibody [JG57-38] (ET7108-18)

Immunogen

Recombinant protein within 1õ”?200 of human prpf8.

Host

Rabbit

Positive Control

A549, 293T, LOVO, SiHa, rat brain tissue, human colon tissue, human prostate cancer tissue, mouse testis tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JG57-38

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

274 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:2,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

PRPF8

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; 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

TISSUE SPECIFICITY

Widely expressed.

SUBCELLULAR LOCATION

Nucleus. Spliceosome.

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.