Glutathione Peroxidase 1 antibody (N-Term)

Catalog No. ABIN5541474

The Mouse Monoclonal anti-Glutathione Peroxidase 1 antibody has been validated for WB, ICC and IF. It is suitable to detect Glutathione Peroxidase 1 in samples from Human.

Quick Overview for Glutathione Peroxidase 1 antibody (N-Term) (ABIN5541474)

Target: Glutathione Peroxidase 1 (GPX1)

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This Glutathione Peroxidase 1 antibody is un-conjugated

Application: Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF)

Clone: 347

Product Details anti-Glutathione Peroxidase 1 Antibody

Binding Specificity: N-Term

Specificity: This antibody reacts with N-terminal eptipe of GPX.

Cross-Reactivity (Details): Does not wor with Mouse (liver, kidney, plasma, NIH/3T3) and Rat (PC12).

Purification: Protein A agarose

Immunogen: Human erythrocyte glutathione peroxidase

Isotype: IgG1

Application Details

Application Notes: Western blot: 10 μg/mL for chemiluminescence detections system. Immunocytochemistry: 1-10 μ/mL. Not recommended for Immunohistochemistry.

Protocol: 1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mM Tris - HCl, pH 7.2, 250 mM NaCl, 0.1 % NP - 40, 2 mM EDTA, 10 % glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12 , 000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the cold Lysis buffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli' s sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 μ L of sample per lane on a 1 - mm - thick SDS - polyacrylamide gel and carry out electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi - dry transfer 97 66 45 30 20 14.4 - system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 2 0 % MeOH). See the manufacture r 's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 o C. 7) Incubate the membrane with primary antibody d iluted with PBS, pH 7.2 containing 1 % skimmed milk as suggest ed in the APPLICATIONS for 1 hour at room temperature. (The concentration of antibody will depend on the conditions.) 8) Wash the membrane with PBS - T [0.05 % Tween - 20 in PBS ] (5 minutes x 6 times). 9) Incubate the membrane with the 1:5,000 HRP - conjugated anti - mouse IgG diluted with 1 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS - T (5 minutes x 6 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriate chemiluminescence reagent for 1 minute. Remove extra reagent from the membrane by dabbing with paper towel, and seal it in plastic wrap. 12) Expose to an X - ray film in a dark room for 5 minutes. Develop the film as usual. The c ondition for exposure and development may vary. (Positive controls for Western blotting Jurkat, Raji, HeLa) Immunocytochemistry 1) Culture the cells in the appropriate condition on a glass slide. (for example, spread 1x 10e4 cells for one slide, then incubate in a CO 2 incubator for one night.) 2) Fixing: a) or b) a) The cells were fixed with methanol at - 20 o C for 2 minutes and with acetone at 4 o C for 5 minutes. b) Fix the cells by immersing the slide in PBS containing 4 % paraformaldehyde for 20 minutes at roomtemperature. The glass slide was washed with PBS 3 times. Immerse the slide in PBS containing 0.1 % Triton X - 100 for 10 minutes at room temperature. 3) The glass slide was washed with PBS 3 times. 4) Cover the cells with 0.2 % BSA in PBS for 10 minutes to minimize non - specific adsorption of the antibodies to the glass slide. 5) Add the primary antibody diluted with PBS as suggest ed in the APPLICATIONS onto the cells and incubate for 30 minutes at room tem p erature (Optimization of antibody concentration or incubation co n dition are recommended if necessary .) 6) The glass slide was washed with PBS 3 times. 7) Add 100 μ L of 1:100 FITC conjugated anti - mouse IgG diluted with PBS onto the cells. Incubate for 30 mi n utes at room temperature. Keep out light by aluminum foil. 8) The glass slide was washed with PBS 3 times. 9) Wipe excess liquid from slide but take care not to touch the cells. Never leave the cells to dry. 10) Promptly add mounting medium onto the slide, then put a cover slip on it.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: PBS containing 50 % clycerol, pH 7.2. No preservative is contained.

Preservative: Azide free

Storage: -20 °C

Storage Comment: Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.

Target Details for Glutathione Peroxidase 1

Target: Glutathione Peroxidase 1 (GPX1)

Alternative Name: glutathione peroxidase 1,gpx1

Background: Glutathione peroxidase (GPX) is a selenoprotein which catalyzes the reduction of a variety of hydroperoxides, including lipid peroxide and hydrogen peroxide, thereby protecting biomembrane and essential cellular components against oxidative damage.

UniProt: P07203

Pathways: Thyroid Hormone Synthesis, Sensory Perception of Sound, Skeletal Muscle Fiber Development, Cell RedoxHomeostasis, Negative Regulation of intrinsic apoptotic Signaling, SARS-CoV-2 Protein Interactome