LONP1 Recombinant Rabbit Monoclonal Antibody [PSH28-77]

Overview

Product Name

LONP1 Recombinant Rabbit Monoclonal Antibody [PSH28-77] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human LONP1 aa 560-959.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC(Intra), IP

Target Molecular Weight

Predicted band size: 106 kDa

Positive Control

HeLa (Human cervical adenocarcinoma cell) cell lysate, HEK-293 (Human embryonic kidney cell) cell lysate, U-87 MG (Human glioblastoma cell) cell lysate, PC-3 (Human prostate cancer cell) cell lysate, Hep G2 (Human hepatocellular carcinoma cell) cell lysate, COS-1 (African green monkey kidney fibroblast) cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate.

Conjugation

unconjugated

Clone Number

PSH28-77

Reactivity Data

Tested

Verified (internally validated)

Published

Reported in literature (not internally validated)

Predicted

Predicted reactive (based on sequence homology)

Not recommended

Not recommended (failed internal validation)

	WB	IHC-P	FC(Intra)	IF-Cell	IP
human
mouse
rat

Product Features

Form

Liquid

Concentration

0.5 mg/mL.(The concentration of this product may be batch-dependent)

Lot Concentration Lookup

Storage Instructions

Store at 2-8℃. Avoid freeze.

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

Target

Function

Lon protease homolog, mitochondrial is a protease, an enzyme that in humans is encoded by the LONP1 gene. Lon protease (LONP1) is a conserved serine peptidase identified from bacteria to eukaryotic cells. In mitochondrial matrix, a majority of damaged proteins is removed via proteolysis led by Lon protease, which is an essential mechanism for mitochondrial protein quality control. LONP1 is the major protease responsible for the recognition and removal of unfolded proteins in the mitochondrial matrix and thereby protects the cell from the accumulation of aggregated proteins in the mitochondrion. However LONP1 is unable to recognise or degrade model aggregated proteins. LONP1 is also a DNA-binding protein that participates in mtDNA maintenance and gene expression regulation. LONP1 degrades mitochondrial transcription factor A (TFAM) when substrate is modified by post-translational modifications (PTMs) such as phosphorylation, regulating mtDNA copy number and metabolism to maintain the TFAM/mtDNA ratio necessary to control replication and transcription.

Background References

1. Liu Y et al. Artemisinins ameliorate polycystic ovarian syndrome by mediating LONP1-CYP11A1 interaction. Science. 2024 Jun

2. Bai M et al. LONP1 targets HMGCS2 to protect mitochondrial function and attenuate chronic kidney disease. EMBO Mol Med. 2023 Feb

Subcellular Location

Mitochondrion matrix.

UNIPROT

SwissProt: P36776 Human

SwissProt: Q8CGK3 Mouse

SwissProt: Q924S5 Rat

Synonyms

Lon protease homolog, mitochondrial

EC:3.4.21.53

LONHs

Lon protease-like protein UniRule annotation (LONP)

Mitochondrial ATP-dependent protease Lon

Mitochondrial chaperone LONP1

Serine protease 15

LONP1

Lon

PRSS15

Images

This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Western blot analysis of LONP1 on different lysates with Rabbit anti-LONP1 antibody (HA724808) at 1/2,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HEK-293 (Human embryonic kidney cell) cell lysate
Lane 3: U-87 MG (Human glioblastoma cell) cell lysate
Lane 4: PC-3 (Human prostate cancer cell) cell lysate
Lane 5: Hep G2 (Human hepatocellular carcinoma cell) cell lysate
Lane 6: COS-1 (African green monkey kidney fibroblast) cell lysate
Lane 7: NIH/3T3 (Mouse fibroblast) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 1 minute 50 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724808, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 106 kDa
Observed band size: 106 kDa
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: SH-SY5Y (Human neuroblastoma cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724808, 1/1,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: The mitochondria dyes was Mito-Tracker (Beyotime, Red). The nuclear counterstain was DAPI (Blue).
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblast)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724808, 1/200, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: The mitochondria dyes was Mito-Tracker (Beyotime, Red). The nuclear counterstain was DAPI (Blue).
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: Rat
Sample: C6 (Rat glioma cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724808, 1/1,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: The mitochondria dyes was Mito-Tracker (Beyotime, Red). The nuclear counterstain was DAPI (Blue).
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Colon
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724808, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Lung
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724808, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: Human
Sample: SH-SY5Y (Human neuroblastoma cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA724808 (1/500, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblasts)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA724808 (1/500, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: Rat
Sample: C6 (Rat glioma cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA724808 (1/500, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Immunoprecipitation (IP)

LONP1 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724808 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724808 at 1/2,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA724808 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724808 in HeLa cell lysate

Exposure time: 10 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724808, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 106 kDa
Observed band size: 106 kDa
This data was developed using HA724808, the same antibody clone in a different buffer formulation.
Immunoprecipitation (IP)

LONP1 was immunoprecipitated in 0.2 mg NIH/3T3 (Mouse fibroblast) cell lysate with HA724808 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724808 at 1/2,000 dilution. HRP Conjugated Rabbit IgG kappa light chain Mouse Monoclonal Antibody(M1208-2) at 1/5000 dilution was used for 1 hour at room temperature.

Lane 1: NIH/3T3 cell lysate (input)
Lane 2: HA724808 IP in NIH/3T3 cell lysate
Lane 3: Rabbit IgG instead of HA724808 in NIH/3T3 cell lysate

Exposure time: 46 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724808, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 106 kDa
Observed band size: 106 kDa