ENO1 Recombinant Rabbit Monoclonal Antibody [PSH26-93]
cat.: HA724658
| 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: | 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: | 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. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC(Intra) IP |
1:5,000 1:50-1:500 1:1,000 1:1,000 1-2μg/sample |
| 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 |
HA724658 - Page 2
Images
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Fig1:
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 |
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Fig2:
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 |
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Fig3:
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 |
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Fig4:
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 |
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Fig5:
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 |
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Fig6:
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 |
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Fig7:
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 |
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Fig8:
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) |
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Fig9:
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). |
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Fig10:
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). |
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Fig11:
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). |
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Fig12:
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) |
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Fig13:
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. |
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Fig14:
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. |
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Fig15:
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. |
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Fig16:
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. |
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Fig17:
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. |
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Fig18:
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. |
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Fig19:
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. |
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Fig20:
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. |
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Fig21:
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. |
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Fig22:
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. |
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Fig23:
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 |
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Fig24:
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”.