ENO1 Recombinant Rabbit Monoclonal Antibody [PSH26-93]
cat.: HA752054
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Green monkey, Pig
Applications: WB, IF-Cell, IHC-P, FC(Intra), IP
Clonality: Monoclonal
Clone number: PSH26-93
Form: Liquid
Storage condition: Store at 2-8℃. Avoid freeze.
Storage buffer: PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 47 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human ENO1 aa 161-210.
Positive control: HeLa (Human cervical adenocarcinoma cell) cell lysate, HEK-293 (Human embryonic kidney cell) cell lysate, SH-SY5Y (Human neuroblastoma cell) cell lysate, HepG2 (Human hepatocellular carcinoma cell) cell lysate, MCF7 (Human breast cancer cell) cell lysate, Jurkat(Human T-lymphoblastic cells) cell lysates, COS-1 (African green monkey kidney fibroblast) cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate, C2C12 (Mouse myoblast) cell lysate, RAW264.7 (Mouse myoblast) cell lysate, C6 (Rat glioma cell) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate, Mouse brain tissue lysate, Mouse kidney tissue lysate, Mouse liver tissue lysate, Mouse pancreas tissue lysate, Rat brain tissue lysate, Rat kidney tissue lysate, Rat heart tissue lysate, Rat liver tissue lysate, Rat pancreas tissue lysate, Pig brain tissue lysate, Pig cerebellum tissue lysate.
Subcellular location: Cytoplasm, Cell membrane, myofibril, sarcomere, M line.
Uniprot #: SwissProt: P06733 Human | P17182 Mouse | P04764 Rat
Alternative names: 2 phospho D glycerate hydro lyase 2-phospho-D-glycerate hydro-lyase Alpha enolase Alpha enolase like 1 Alpha-enolase C myc promoter binding protein C-myc promoter-binding protein EC 4.2.1.11 eno1 ENO1L1 ENOA_HUMAN Enolase 1 (alpha) Enolase 1 (alpha) like 1 Enolase 1 Enolase alpha MBP 1 MBP-1 MBP1 MBPB1 MPB 1 MPB-1 MPB1 MYC promoter binding protein 1 NNE Non neural enolase Non-neural enolase Phosphopyruvate hydratase Plasminogen binding protein Plasminogen-binding protein PPH Tau crystallin
HA752054 - Page 2
Images
HA752054_1.jpg Fig1: This data was developed using HA752054, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA752054) 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: HA752054, 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
HA752054_2.jpg Fig2: This data was developed using HA752054, 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 (HA752054) 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: HA752054, 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
HA752054 - Page 3
HA752054_3.jpg Fig3: This data was developed using HA752054, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA752054) 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: HA752054, 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
HA752054_4.jpg Fig4: This data was developed using HA752054, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA752054) 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: HA752054, 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
HA752054 - Page 4
HA752054_5.jpg Fig5: This data was developed using HA752054, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA752054) 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: HA752054, 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
HA752054_6.jpg Fig6: This data was developed using HA752054, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA752054) 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: HA752054, 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
HA752054 - Page 5
HA752054_7.jpg Fig7: This data was developed using HA752054, the same antibody clone in a different buffer formulation.
Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (HA752054) 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: HA752054, 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
HA752054_8.jpg Fig8: This data was developed using HA752054, 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: HA752054, 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)
HA752054 - Page 6
HA752054_9.jpg Fig9: This data was developed using HA752054, 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: HA752054, 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).
HA752054_10.jpg Fig10: This data was developed using HA752054, 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: HA752054, 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).
HA752054 - Page 7
HA752054_11.jpg Fig11: This data was developed using HA752054, 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: HA752054, 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).
HA752054_12.jpg Fig12: This data was developed using HA752054, 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: HA752054, 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)
HA752054 - Page 8
HA752054_13.jpg Fig13: This data was developed using HA752054, 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: HA752054, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA752054_14.jpg Fig14: This data was developed using HA752054, 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: HA752054, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA752054 - Page 9
HA752054_15.jpg Fig15: This data was developed using HA752054, 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: HA752054, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA752054_16.jpg Fig16: This data was developed using HA752054, 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: HA752054, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA752054 - Page 10
HA752054_17.jpg Fig17: This data was developed using HA752054, 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: HA752054, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA752054_18.jpg Fig18: This data was developed using HA752054, 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: HA752054 (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.

HA752054 - Page 11
HA752054_19.jpg Fig19: This data was developed using HA752054, 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: HA752054 (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.
HA752054_20.jpg Fig20: This data was developed using HA752054, 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: HA752054 (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.

HA752054 - Page 12
HA752054_21.jpg Fig21: This data was developed using HA752054, 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: HA752054 (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.
HA752054_22.jpg Fig22: This data was developed using HA752054, 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: HA752054 (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.

HA752054 - Page 13
HA752054_23.JPG Fig23: This data was developed using HA752054, 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 HA752054 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA752054 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: HA752054 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA752054 in HeLa cell lysate

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

Primary dilution: HA752054, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 47 kDa
Observed band size: 47 kDa
HA752054_24.jpg Fig24: This data was developed using HA752054, 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.