TPP1 Recombinant Rabbit Monoclonal Antibody [PSH18-09]
Catalog# HA723972
TPP1 Recombinant Rabbit Monoclonal Antibody [PSH18-09]
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WB
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IHC-P
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IF-Cell
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FC
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IP
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Human
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Mouse
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Rat
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HA751655
不含抗保成分
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unconjugated
Overview
Product Name
TPP1 Recombinant Rabbit Monoclonal Antibody [PSH18-09]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human TPP1 aa 1-563.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell, FC, IP
Molecular Weight
Predicted band size: 61 kDa
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.
Conjugation
unconjugated
Clone Number
PSH18-09
Reactivity Data
| WB | IHC-P | IF-Cell | FC | IP | |
|---|---|---|---|---|---|
| Human |
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| Mouse |
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| Rat |
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Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IHC-P
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1:1,000
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IF-Cell
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1:100-1:200
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FC
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1:1,000
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IP
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1-2μg/sample
Target
Function
This gene encodes a member of the sedolisin family of serine proteases. The protease functions in the lysosome to cleave N-terminal tripeptides from substrates, and has weaker endopeptidase activity. It is synthesized as a catalytically-inactive enzyme which is activated and auto-proteolyzed upon acidification. Mutations in this gene result in late-infantile neuronal ceroid lipofuscinosis, which is associated with the failure to degrade specific neuropeptides and a subunit of ATP synthase in the lysosome.
Background References
1. Lin L, Sohar I, Lackland H, Lobel P. The human CLN2 protein/tripeptidyl-peptidase I is a serine protease that autoactivates at acidic pH. J Biol Chem. 2001 Jan 19;276(3):2249-55.
2. Guhaniyogi J, Sohar I, Das K, Stock AM, Lobel P. Crystal structure and autoactivation pathway of the precursor form of human tripeptidyl-peptidase 1, the enzyme deficient in late infantile ceroid lipofuscinosis. J Biol Chem. 2009 Feb 6;284(6):3985-97.
Subcellular Location
Lysosome, Melanosome.
Synonyms
Cell growth inhibiting gene 1 protein antibody
Cell growth-inhibiting gene 1 protein antibody
Ceroid lipofuscinosis neuronal 2 antibody
Ceroid lipofuscinosis neuronal 2 late infantile (Jansky Bielschowsky disease) antibody
Ceroid lipofuscinosis neuronal 2 late infantile antibody
CLN 2 antibody
CLN2 antibody
GIG 1 antibody
GIG1 antibody
Growth inhibiting protein 1 antibody
ExpandImages
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Western blot analysis of TPP1 on different lysates with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Western blot analysis of TPP1 on different lysates with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunohistochemical analysis of paraffin-embedded mouse placenta tissue with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunohistochemical analysis of paraffin-embedded rat placenta tissue with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunocytochemistry analysis of Raji cells labeling TPP1 with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Immunocytochemistry analysis of HepG2 cells labeling TPP1 with Rabbit anti-TPP1 antibody (HA723972) 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 (HA723972) 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. -
Flow cytometric analysis of Raji cells labeling TPP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723972, 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). -
Flow cytometric analysis of HepG2 cells labeling TPP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723972, 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). -
TPP1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA723972 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723972 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: HA723972 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA723972 in HeLa cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 6 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"