SOD1 antibody (AA 2-154)

Catalog No. ABIN2913314

Product Details anti-SOD1 Antibody

Target: SOD1 (Superoxide Dismutase 1, Soluble (SOD1))

Binding Specificity: AA 2-154

Reactivity: Human, Horse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This SOD1 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunofluorescence (fixed cells) (IF/ICC)

Purification: Antigen-specific affinity chromatography followed by Protein A affinity chromatography

Immunogen: SOD1 (Ala2-Pro154)

Isotype: IgG

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (WB)

Validation #100074 (Western Blotting)

by: Royal Veterinary College, London

No.: #100074

Date: 11/17/2016

Antigen: SOD-1

Method validated: Western Blotting

Positive Control: Horse Fibroblasts, human SH-SY5Y

Notes: ABIN3058918 successfully labels SOD-1 in lysates of equine fibroblast and human SH-SY5Y cells.

Validation Images

Total protein lysates from horse fibroblasts and human SH-SY5Y cells were separated were separated on a polyacrylamide gel under denaturing conditions. SOD-1 and TUBB protein bands were revealed as described in the protocol section.

Full Methods

Primary Antibody: ABIN3058918

Secondary Antibody: Goat anti-rabbit-IgG HRP-linked whole antibody (Dako, D0487)

Full Protocol:

• Seed 2x10⁵ horse fibroblasts in a T25 flask and grow for 48h at 5% CO₂ in supplemented with Dulbecco’s modified Eagle media (Sigma-Aldrich, D5546), with 10% heat-inactivated fetal calf serum (FCS) and 2mM (1%) L-glutamine and 1% penicillin (10IU/ml)/streptomycin (100µg/ml) to form a monolayer of 70-90%.
• Human SH-SY5Y are cultured in identical conditions.
• Wash cells once ice cold PBS.
• Add 500µl chilled RIPA buffer (50mM Tris pH 7.5, 150mM NaCl, 1% NP40, 0.5% Na-deoxychlorate, 0.5% SDS, 1mM EDTA, 1mM EGTA, 1mM PMSF and 1x protease inhibitor) to each flask and incubate for 1min on ice.
• Scrape each monolayer and transfer protein aggregates to a separate 1.5ml microcentrifuge tube.
• Incubate for 20min on ice.
• Centrifuge at 14000rpm for 20min at 4°C.
• Remove the supernatant and determine the concentration of soluble proteins against a BSA protein standard using a DC protein assay (Biorad, 5000112).
• Adjust protein samples to 0.3mg/ml total protein, using RIPA buffer and loading buffer.
• After boiling for 5-10min, 1-10µg of each sample is separated on an Any kD Mini-PROTEAN TGX Stain-Free Precast Gel (Biorad, 4568123) at 100V for 90min.
• Transfer proteins onto a PVDF blotting membrane (Hybond P, GE Healthcare Life Sciences, RPN303D) for 90min at 300mA.
• Block the membrane with PBST containing 10% dry milk for 1h at RT.
• Incubation with primary SOD-1 antibody (antibodies-online, ABIN3058918) diluted 1:1000 and a TUBB loading control antibody (abcam, ab6046) diluted 1:1000 in PBST containing 5% dry milk ON at 4°C.
• Wash 5 times with PBST over 1 hour whilst shaking.
• Incubation with secondary goat anti-rabbit-IgG horseradish peroxidase-linked whole antibody (Dako, D0487) diluted 1:5000 in PBST for 1h at RT.
• Wash 5 times with PBST over 1 hour whilst shaking.
• Blots were developed with Amersham ECL Prime Western Blotting Detection reagent (GE Healthcare Life Sciences, RPN2232) and visualized on a Kodak developer.

Experimental Notes: The same human cell lysates were also used for a different blot with an anti-human SOD-1 antibody (lab stock). A band of the same molecular weight as in the horse fibroblast lysates was revealed.

Independent Validation (ICC)

Validation #100075 (Immunocytochemistry)

by: Royal Veterinary College, London

No.: #100075

Date: 11/17/2016

Antigen: SOD-1

Method validated: Immunocytochemistry

Positive Control: Horse Fibroblasts

Notes: ABIN3058918 achieves SOD-1 staining in horse fibroblasts when grown in a monolayer.

Validation Images

Horse fibroblasts were grown in a monolayer and stained with ABIN3058918 (red) as desecribed in the protocol. DAPI (blue) was used to visualize nuclei.

Full Methods

Primary Antibody: ABIN3058918

Secondary Antibody: anti-rabbit labeled with Alexa Fluor 594

Full Protocol:

• Horse fibroblasts are seeded onto glass coverslips pre-treated with 1:10 diluted Matrigel (BD Biosciences, 354234) at a density of 1x10⁵ cells per coverslip in a 24 well plate.
• Incubate for 48h at 37°C in 5% CO₂.
• Wash coverslips with PBS.
• Fix coverslips with 4% paraformaldehyde (PFA) and 0.1% triton for 20min at 4°C, followed by 8% PFA for 30min at RT.
• Permeabilize cells with 0.5% triton for 30min at RT.
• Wash coverslips 3 times with PBS.
• Incubate coverslips with primary rabbit anti SOD-1 antibody (antibodies-online, ABIN3058918) diluted 1:40 in a dark, humidified chamber for 1h at RT. A negative control was incubated without primary antibody.
• Wash coverslips 3 times with PBS.
• Incubate with secondary goat anti rabbit labeled with Alexa Fluor 594 (ThermoFisher Scientific, A10239) diluted 1:500 for 1h at RT.
• Wash coverslips 3 times with PBS.
• Mount coverslips in a hard setting mount with VECTASHIELD HardSet Antifade Mounting Medium with DAPI (Vector Laboratories, H-1500).
• Fluorescence microscopy is then used to image each coverslip.

Handling

Format: Liquid

Concentration: 500 μg/mL

Buffer: 0.01M PBS,  pH 7.4, containing 0.05 % Proclin-300, 50 % glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4 °C for frequent use. Aliquot and store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

References for anti-SOD1 antibody (ABIN2913314)

Kronstein-Wiedemann, Stadtmüller, Traikov, Georgi, Teichert, Yosef, Wallenborn, Karl, Schütze, Wagner, El-Armouche, Tonn: "SARS-CoV-2 Infects Red Blood Cell Progenitors and Dysregulates Hemoglobin and Iron Metabolism." in: Stem cell reviews and reports, (2022) (PubMed).

Target Details for SOD1

Target: SOD1 (Superoxide Dismutase 1, Soluble (SOD1))

Alternative Name: Superoxide Dismutase 1 (SOD1 Products)

Synonyms: LOC692639 antibody, ALS antibody, ALS1 antibody, IPOA antibody, SOD antibody, hSod1 antibody, homodimer antibody, CG11793 antibody, Cu antibody, Cu-Zn SOD antibody, Cu/Zn SOD antibody, Cu/Zn sod antibody, Cu/Zn superoxide dismutase antibody, Cu/ZnSOD antibody, CuSOD antibody, CuZn SOD antibody, CuZn-SOD antibody, CuZn-SOD1 antibody, CuZnSOD antibody, Cu[2+]/Zn[2+]SOD antibody, Dmel\\CG11793 antibody, G antibody, Mn SOD antibody, SOD-1 antibody, SOD1 antibody, Sod-1 antibody, Sod1 antibody, To antibody, To-1 antibody, Zn SOD antibody, Zn Sod antibody, Zn-SOD antibody, ZnSod antibody, cSOD antibody, cSod antibody, dSOD1 antibody, l(3)108 antibody, l(3)68Af' antibody, l(3)G antibody, sod antibody, sod1 antibody, CU/ZN-SOD antibody, SODC antibody, DKFZP469M1833 antibody, B430204E11Rik antibody, Cu/Zn-SOD antibody, Ipo-1 antibody, Ipo1 antibody, SOD1L1 antibody, XSODB antibody, als antibody, als1 antibody, ipoa antibody, sod1-a antibody, ZSOD antibody, cuzn antibody, Cu/Zn superoxide dismutase antibody, superoxide dismutase 1 antibody, Superoxide dismutase 1 antibody, superoxide dismutase 1, soluble antibody, superoxide dismutase 1 S homeolog antibody, Superoxide dismutase [Cu-Zn] antibody, superoxide dismutase 1 L homeolog antibody, superoxide dismutase [Cu-Zn]-like antibody, superoxide dismutase [Cu-Zn] antibody, superoxide dismutase Sod1 antibody, SOD antibody, SOD1 antibody, Sod1 antibody, sod1 antibody, sod1.S antibody, sod-1 antibody, sod1.L antibody, LOC101451855 antibody, LOC101115136 antibody

Pathways: Sensory Perception of Sound, Transition Metal Ion Homeostasis