HINT1 Antibody

• Catalog Number: orb1926327
• Category: Antibodies
• Description: Purified Mouse Monoclonal Antibody (Mab)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 1500CT836.13.93
• Tested applications: FC, IF, IHC-P, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG1,k
• Dilution range: IF: 1:25, WB: 1:4000, IHC-P: 1:25, FC: 1:25
• Form/Appearance: Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein G column, followed by dialysis against PBS.
• Conjugation: Unconjugated
• MW: 13802 Da
• Target: This HINT1 antibody is generated from a mouse immunized with a recombinant protein of human HINT1.
• UniProt ID: P49773
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles
• Alternative names: Histidine triad nucleotide-binding protein 1, 3---
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

All lanes: Anti-HINT1 Antibody at 1:4000 dilution. Lane 1: Jurkat whole cell lysates. Lane 2: Raji whole cell lysates. Lane 3: HepG2 whole cell lysates. Lane 4: rat brain lysates. Lane 5: mouse liver lysates. Lysates/proteins at 20 μg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 14 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Staining HINT1 in human kidney sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma cell line) cells labeling HINT1 at 1/25 dilution, followed by Dylight 488-conjugated goat anti-mouse IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on U-2 OS cell line. Cytoplasmic actin is detected with Dylight 554 Phalloidin at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).

Overlay histogram showing Jurkat cells stained (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37°C. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight 488 Conjugated Highly Cross-Adsorbed at 1/400 dilution for 40 min at 37°C. Isotype control antibody (blue line) was mouse IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of > 10000 events was performed.