PP-X Rabbit Polyclonal Antibody
cat.: HA500276
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, FC, IF-Cell
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 35 kDa
Isotype: IgG
Immunogen: Recombinant protein within human PP-X aa 151-307 / 307.
Positive control: HeLa cell lysate, MCF7 cell lysate, RAW264.7 cell lysate, Rat kidney tissue lysate, Rat brain tissue lysate, human colon tissue, HeLa, NIH/3T3.
Subcellular location: Cytoplasm, Nucleus, cytoskeleton, microtubule organizing center, centrosome.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IHC-P
1:5,000
1:50-1:100
1:1,000
1:100-1:500
Uniprot #: SwissProt: P60510 Human | P97470 Mouse | Q5BJ92 Rat
Alternative names: PP X PP-X Pp4 PP4C PP4C_HUMAN PPH3 PPP4 ppp4c PPX protein phosphatase 4 (formerly X), catalytic subunit Protein phosphatase 4 catalytic subunit Protein phosphatase X protein phosphatase X, catalytic subunit Serine/threonine protein phosphatase 4 catalytic subunit Serine/threonine-protein phosphatase 4 catalytic subunit
Images
HA500276_1.jpg Fig1: Western blot analysis of PP-X on different lysates with Rabbit anti-PP-X antibody (HA500276) at 1/5,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: MCF7 cell lysate (20 µg/Lane)
Lane 3: RAW264.7 cell lysate (20 µg/Lane)
Lane 4: Rat kidney tissue lysate (40 µg/Lane)
Lane 5: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 35 kDa
Observed band size: 35 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 (HA500276) 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.
HA500276_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling PP-X with Rabbit anti-PP-X antibody (HA500276) at 1/50 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-PP-X antibody (HA500276) at 1/50 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.
HA500276_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling PP-X with Rabbit anti-PP-X antibody (HA500276) 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-PP-X antibody (HA500276) 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.
HA500276_4.jpg Fig4: Flow cytometric analysis of HeLa cells labeling PP-X.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500276, 1/1,000) (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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.