PNKP Recombinant Rabbit Monoclonal Antibody [JE64-21]
cat.: HA721044
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: JE64-21
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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: 57 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human PNKP aa 51-100 / 521.
Positive control: Jurkat cell lysate, HepG2 cell lysate, PC-3 cell lysate, Hela cell lysate, human breast carcinoma tissue, human kidney tissue, Hela.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P

1:500-1,000
1:100
1:400
Uniprot #: SwissProt: Q96T60 Human
Alternative names: 2''(3'')-polynucleotidase 2'(3')-polynucleotidase Bifunctional polynucleotide phosphatase/kinase DEM 1 DEM1 DNA 5' kinase/3' phosphatase DNA 5''-kinase/3''-phosphatase EIEE10 Homo sapiens polynucleotide kinase 3' phosphatase (PNKP) MCSZ PNK 1 PNK1 Pnkp PNKP DNA kinase PNKP_HUMAN Polynucleotide 3'-phosphatase Polynucleotide 5' hydroxyl kinase Polynucleotide 5''-hydroxyl-kinase Polynucleotide kinase 3 prime phosphatase Polynucleotide kinase 3' phosphatase Polynucleotide Kinase Polynucleotide kinase-3''-phosphatase
Images
HA721044_1.jpg Fig1: Western blot analysis of PNKP on different lysates with Rabbit anti-PNKP antibody (HA721044) at 1/1,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: HepG2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 57 kDa
Observed band size: 60 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 (HA721044) at 1/1,000 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.
HA721044_2.jpg Fig2: Western blot analysis of PNKP on different lysates with Rabbit anti-PNKP antibody (HA721044) at 1/500 dilution.

Lane 1: PC-3 cell lysate
Lane 2: Hela cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 57 kDa
Observed band size: 60 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 (HA721044) 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.
HA721044_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-PNKP antibody (HA721044) 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 (HA721044) 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.
HA721044_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PNKP antibody (HA721044) 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 (HA721044) 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.
HA721044_5.jpg Fig5: Immunocytochemistry analysis of Hela cells labeling PNKP with Rabbit anti-PNKP antibody (HA721044) at 1/100 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-PNKP antibody (HA721044) at 1/100 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.