PML Protein Rabbit Polyclonal Antibody
cat.: HA500540
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 98 kDa
Isotype: IgG
Immunogen: Recombinant protein within human PML Protein aa 1-600 / 882.
Positive control: HeLa cell lysate, HEK-293 cell lysate, K-562 cell lysate, A549 cell lysate, MDA-MB-231 cell lysate, A431 cell lysate, human kidney tissue lysate, MDA-MB-231, A431, human brain tissue, human breast tissue, human breast cancer tissue, human testis tissue, mouse breast tissue, rat breast tissue.
Subcellular location: Nucleus, Cytoplasm, PML body, nucleolus, Endoplasmic reticulum membrane, Early endosome membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:2,000
1:100
1:1,000
1:1,000
Uniprot #: SwissProt: P29590 Human | Q60953 Mouse
Entrez Gene: 315713 Rat
Alternative names: Acure promyelocytic leukemia, inducer of MYL Pml PML_HUMAN PP8675 Probable transcription factor PML Promyelocytic leukemia Promyelocytic leukemia inducer of Promyelocytic leukemia protein Protein PML RING finger protein 71 RNF 71 RNF71 TRIM 19 Tripartite motif protein TRIM19 Tripartite motif-containing protein 19
Images
HA500540_1.jpg Fig1: Western blot analysis of PML Protein on different lysates with Rabbit anti-PML Protein antibody (HA500540) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: K-562 cell lysate
Lane 4: A549 cell lysate
Lane 5: MDA-MB-231 cell lysate
Lane 6: A431 cell lysate
Lane 7: Human kidney tissue lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 98 kDa
Observed band size: 50-130 kDa

Exposure time: 43 seconds;

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 (HA500540) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA500540_2.jpg Fig2: Immunocytochemistry analysis of MDA-MB-231 cells labeling PML Protein with Rabbit anti-PML Protein antibody (HA500540) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-PML Protein antibody (HA500540) at 1/500 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 (M1305-2, 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.
HA500540_3.jpg Fig3: Immunocytochemistry analysis of A431 cells labeling PML Protein with Rabbit anti-PML Protein antibody (HA500540) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-PML Protein antibody (HA500540) at 1/250 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 (M1305-2, 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.
HA500540_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PML Protein antibody (HA500540) at 1/2,000 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 (HA500540) at 1/2,000 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.
HA500540_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-PML Protein antibody (HA500540) at 1/2,000 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 (HA500540) at 1/2,000 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.
HA500540_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-PML Protein antibody (HA500540) at 1/2,000 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 (HA500540) at 1/2,000 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.
HA500540_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PML Protein antibody (HA500540) at 1/2,000 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 (HA500540) at 1/2,000 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.
HA500540_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse breast tissue with Rabbit anti-PML Protein antibody (HA500540) at 1/500 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 (HA500540) at 1/500 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.
HA500540_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat breast tissue with Rabbit anti-PML Protein antibody (HA500540) at 1/500 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 (HA500540) at 1/500 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.
HA500540_10.jpg Fig10: Flow cytometric analysis of MDA-MB-231 cells labeling PML Protein.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500540, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA500540_11.jpg Fig11: Flow cytometric analysis of A431 cells labeling PML Protein.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500540, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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