PRPF4 Recombinant Rabbit Monoclonal Antibody [JE65-37]
cat.: HA721063
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: JE65-37
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 58 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human PRPF4 aa 1-50/522.
Positive control: RAW264.7 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, HT-29 cell lysate, SW480 cell lysate, MCF-7 cell lysate, H1395, SiHa, SW480, NIH/3T3, C6, human cervical carcinoma tissue, human colon tissue.
Subcellular location: Nucleus, Nucleus speckle.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:500-1:5,000
1:50-1:200
1:100-1:400
Uniprot #: SwissProt: O43172 Human | Q9DAW6 Mouse
Entrez Gene: 298095 Rat
Alternative names: hPrp4 HPRP4P PRP4 PRP4 homolog PRP4 pre mRNA processing factor 4 homolog (yeast) PRP4/STK/WD splicing factor PRP4_HUMAN Prp4p PRPF4 RP70 SNRNP60 U4/U6 small nuclear ribonucleoprotein Prp4 U4/U6 snRNP 60 kDa protein WD splicing factor Prp4
Images
HA721063_1.jpg Fig1: Western blot analysis of PRPF4 on different lysates with Rabbit anti-PRPF4 antibody (HA721063) at 1/5,000 dilution.

Lane 1: RAW264.7 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 58 kDa
Observed band size: 58 kDa

Exposure time: 20 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721063) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA721063_2.jpg Fig2: Western blot analysis of PRPF4 on different lysates with Rabbit anti-PRPF4 antibody (HA721063) at 1/500 dilution.

Lane 1: HT-29 cell lysate
Lane 2: SW480 cell lysate
Lane 3: MCF-7 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 58 kDa
Observed band size: 58 kDa

Exposure time: 2 minutes;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721063) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
HA721063_3.jpg Fig3: Immunocytochemistry analysis of H1395 cells labeling PRPF4 with Rabbit anti-PRPF4 antibody (HA721063) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PRPF4 antibody (HA721063) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (Alexa Fluor® 555) were used as the secondary antibody at 1/1,000 dilution.
HA721063_4.jpg Fig4: Immunocytochemistry analysis of SiHa cells labeling PRPF4 with Rabbit anti-PRPF4 antibody (HA721063) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PRPF4 antibody (HA721063) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
HA721063_5.jpg Fig5: Immunocytochemistry analysis of SW480 cells labeling PRPF4 with Rabbit anti-PRPF4 antibody (HA721063) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PRPF4 antibody (HA721063) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721063_6.jpg Fig6: Immunocytochemistry analysis of NIH/3T3 cells labeling PRPF4 with Rabbit anti-PRPF4 antibody (HA721063) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PRPF4 antibody (HA721063) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721063_7.jpg Fig7: Immunocytochemistry analysis of C6 cells labeling PRPF4 with Rabbit anti-PRPF4 antibody (HA721063) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PRPF4 antibody (HA721063) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721063_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue with Rabbit anti-PRPF4 antibody (HA721063) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721063) at 1/400 dilution for 1 hour 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.
HA721063_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PRPF4 antibody (HA721063) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721063) at 1/100 dilution for 1 hour 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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