TPP1 Recombinant Rabbit Monoclonal Antibody [PSH18-09]
cat.: HA751655
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: PSH18-09
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 61 kDa
Isotype: IgG
Immunogen: Recombinant protein within human TPP1 aa 1-563.
Positive control: HepG2 cell lysate, HeLa cell lysate, Jurkat cell lysate, Raji cell lysate, 786-0 cell lysate, A431 cell lysate, JAR cell lysate, U-87 MG cell lysate, Mouse heart tissue lysate, Mouse cerebellum tissue lysate, Mouse skeletal muscle tissue lysate, Rat heart tissue lysate, Rat cerebellum tissue lysate, Rat skeletal muscle tissue lysate, human skeletal muscle tissue, human cerebellum tissue, human placenta tissue, mouse cerebellum tissue, mouse placenta tissue, rat cerebellum tissue, rat placenta tissue, Raji, HepG2.
Subcellular location: Lysosome, Melanosome.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
  IP
1:5,000
1:1,000
1:100-1:200
1:1,000
1-2μg/sample
Uniprot #: SwissProt: O14773 Human | O89023 Mouse | Q9EQV6 Rat
Alternative names: Cell growth inhibiting gene 1 protein Cell growth-inhibiting gene 1 protein Ceroid lipofuscinosis neuronal 2 Ceroid lipofuscinosis neuronal 2 late infantile (Jansky Bielschowsky disease) Ceroid lipofuscinosis neuronal 2 late infantile CLN 2 CLN2 GIG 1 GIG1 Growth inhibiting protein 1 LPIC Lysosomal pepstatin insensitive protease Lysosomal pepstatin-insensitive protease MGC21297 TPP 1 TPP I TPP-1 TPP-I Tpp1 TPP1_HUMAN TPPI Tripeptidyl aminopeptidase Tripeptidyl peptidase I Tripeptidyl-peptidase 1 Tripeptidyl-peptidase I
Images
HA751655_1.jpg Fig1: Western blot analysis of TPP1 on different lysates with Rabbit anti-TPP1 antibody (HA751655) at 1/5,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: Jurkat cell lysate (20 µg/Lane)
Lane 4: Raji cell lysate (20 µg/Lane)
Lane 5: 786-0 cell lysate (20 µg/Lane)
Lane 6: A431 cell lysate (20 µg/Lane)
Lane 7: JAR cell lysate (20 µg/Lane)
Lane 8: U-87 MG cell lysate (20 µg/Lane)

Predicted band size: 61 kDa
Observed band size: 48/61 kDa

Exposure time: 6 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 (HA751655) 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.
HA751655_2.jpg Fig2: Western blot analysis of TPP1 on different lysates with Rabbit anti-TPP1 antibody (HA751655) at 1/5,000 dilution.

Lane 1: Mouse heart tissue lysate (20 µg/Lane)
Lane 2: Mouse cerebellum tissue lysate (20 µg/Lane)
Lane 3: Mouse skeletal muscle tissue lysate (20 µg/Lane)
Lane 4: Rat heart tissue lysate (20 µg/Lane)
Lane 5: Rat cerebellum tissue lysate (20 µg/Lane)
Lane 6: Rat skeletal muscle tissue lysate (20 µg/Lane)

Predicted band size: 61 kDa
Observed band size: 48/61 kDa

Exposure time: 10 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 (HA751655) 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.
HA751655_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-TPP1 antibody (HA751655) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751655) at 1/1,000 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.
HA751655_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-TPP1 antibody (HA751655) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751655) at 1/1,000 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.
HA751655_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-TPP1 antibody (HA751655) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751655) at 1/1,000 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.
HA751655_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-TPP1 antibody (HA751655) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751655) at 1/1,000 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.
HA751655_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse placenta tissue with Rabbit anti-TPP1 antibody (HA751655) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751655) at 1/1,000 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.
HA751655_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-TPP1 antibody (HA751655) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751655) at 1/1,000 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.
HA751655_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat placenta tissue with Rabbit anti-TPP1 antibody (HA751655) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA751655) at 1/1,000 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.
HA751655_10.jpg Fig10: Immunocytochemistry analysis of Raji cells labeling TPP1 with Rabbit anti-TPP1 antibody (HA751655) 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-TPP1 antibody (HA751655) 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.
HA751655_11.jpg Fig11: Immunocytochemistry analysis of HepG2 cells labeling TPP1 with Rabbit anti-TPP1 antibody (HA751655) at 1/200 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-TPP1 antibody (HA751655) at 1/200 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.
HA751655_12.jpg Fig12: Flow cytometric analysis of Raji cells labeling TPP1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751655, 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).
HA751655_13.jpg Fig13: Flow cytometric analysis of HepG2 cells labeling TPP1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751655, 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).
HA751655_14.jpg Fig14: TPP1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751655 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751655 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA751655 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751655 in HeLa cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 6 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.