LONP1 Recombinant Rabbit Monoclonal Antibody [PSH28-77]
cat.: HA724808
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC(Intra), IP
Clonality: Monoclonal
Clone number: PSH28-77
Form: Liquid
Storage condition: 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.
Concentration: 0.5ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 106 kDa
Isotype: IgG
Immunogen: Recombinant protein within human LONP1 aa 560-959.
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.
Subcellular location: Mitochondrion matrix.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC(Intra)
  IP
1:2,000
1:200-1:1,000
1:500
1:500
1-2μg/sample
Uniprot #: SwissProt: P36776 Human | Q8CGK3 Mouse | Q924S5 Rat
Alternative names: 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
HA724808_1.jpg Fig1: 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
HA724808_2.jpg Fig2: 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).
HA724808_3.jpg Fig3: 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).
HA724808_4.jpg Fig4: 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).
HA724808_5.jpg Fig5: 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.
HA724808_6.jpg Fig6: 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.
HA724808_7.jpg Fig7: 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.

HA724808_8.jpg Fig8: 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.

HA724808_9.jpg Fig9: 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.

HA724808_10.jpg Fig10: 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
HA724808_11.jpg Fig11: 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
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.