ENO1 Recombinant Rabbit Monoclonal Antibody [PSH26-93] - BSA and Azide free

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA724658) at 1/5,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HEK-293 (Human embryonic kidney cell) cell lysate
Lane 3: SH-SY5Y (Human neuroblastoma cell) cell lysate
Lane 4: HepG2 (Human hepatocellular carcinoma cell) cell lysate
Lane 5: MCF7 (Human breast cancer cell) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 16 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724658, 1/5,000 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: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on Jurkat(Human T-lymphoblastic cells) cell lysate with Rabbit anti-ENO1 antibody (HA724658) at 1/5,000 dilution.

Lysates/proteins at 15 µg/Lane.
Exposure time: 16 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724658, 1/5,000 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: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA724658) at 1/5,000 dilution.

Lane 1: COS-1 (African green monkey kidney fibroblast) cell lysate
Lane 2: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 3: C2C12 (Mouse myoblast) cell lysate
Lane 4: RAW264.7 (Mouse myoblast) cell lysate
Lane 5: C6 (Rat glioma cell) cell lysate
Lane 6: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 16 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724658, 1/5,000 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: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA724658) at 1/5,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Mouse kidney tissue lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 16 seconds ; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724658, 1/5,000 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: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA724658) at 1/5,000 dilution.

Lane 1: Mouse liver tissue lysate
Lane 2: Mouse pancreas tissue lysate
Lane 3: Rat brain tissue lysate
Lane 4: Rat kidney tissue lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 16 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724658, 1/5,000 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: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA724658) at 1/5,000 dilution.

Lane 1: Rat heart tissue lysate
Lane 2: Rat liver tissue lysate
Lane 3: Rat pancreas tissue lysate
Lane 4: Pig brain tissue lysate
Lane 5: Pig cerebellum tissue lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 16 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724658, 1/5,000 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: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA724658) at 1/5,000 dilution.

Lane 1: His-tagged Human ENO1 recombinant protein
Lane 2: His-tagged Human ENO2 recombinant protein
Lane 3: His-tagged Human ENO3 recombinant protein

Lysates/proteins at 20 ng/Lane.
Exposure time: 25 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724658, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial 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: HA724658, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

Subcellular localization consistent with literature. (PMID: 28525970)

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: RAW264.7 (Mouse monocytic macrophage leukemia 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: HA724658, 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

This data was developed using HA724658, 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: HA724658, 1/50, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: Rat
Sample: PC-12 (Rat pheochromocytoma cell (undifferentiated))

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: HA724658, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunocytochemistry (IF-cell)

Species: African green monkey
Sample: COS-1 (African green monkey kidney 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: HA724658, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

Subcellular localization consistent with literature. (PMID: 28525970)

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Brain
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: HA724658, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Pancreas
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: HA724658, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Brain
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: HA724658, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Pancreas
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: HA724658, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Brain
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: HA724658, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

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: HA724658 (1/1,000, 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 HA724658, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: Mouse
Sample: RAW264.7 (Mouse monocytic macrophage leukemia cell)

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: HA724658 (1/1,000, 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 HA724658, 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: HA724658 (1/1,000, 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 HA724658, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: Rat
Sample: PC-12 (Rat pheochromocytoma cells (undifferentiated))

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: HA724658 (1/1,000, 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 HA724658, the same antibody clone in a different buffer formulation.
Application: Flow Cytometry (Intra)

Species: African green monkey
Sample: COS-1 (African green monkey kidney fibroblast)

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: HA724658 (1/1,000, 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 HA724658, the same antibody clone in a different buffer formulation.
Immunoprecipitation (IP)

ENO1 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724658 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724658 at 1/5,000 dilution. HRP Conjugated Rabbit IgG kappa light chain Mouse Monoclonal Antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA724658 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724658 in HeLa cell lysate

Exposure time: 2 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724658, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 47 kDa
Observed band size: 47 kDa

This data was developed using HA724658, the same antibody clone in a different buffer formulation.
Application: Enzyme-Linked Immunosorbent Assay (ELISA)

HA724658 can only recognize ENO1 peptide, and does not recognize ENO2 and ENO3 peptides.