Actin anticorps

N° du produit ABIN3181106

Détail du produit anti-Actin anticorps

Antigène: Actin

Reactivité: Plantes

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp Actin est non-conjugé

Application: Western Blotting (WB)

Specificité: The antibody detects endogenous plant actin protein.

Attributs du produit: Mouse Monoclonal to Plant actin.

Purification: The antibody was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen.

Clone: Q30

Isotype: IgG1

Information d'application

Indications d'application: WB 1:2000-5000

Restrictions: For Research Use only

Independent Validation (WB)

Validation #102343 (Western Blotting)

by: Department of Horticultural genetics and Biotechnology, Mediterranean Agronomic Institute of Chania

No.: #102343

Date: 19.12.2017

Antigène: Actin

Numéro du lot: B3401

Application validée: Western Blotting

Contrôle positif: protein extraction from olive root tissue and tomato flower tissue

Conclusion:

Passed. ABIN3181106 specifically recognizes the actin in both tomato flower and olive roots samples, providing a clear band at the detection with the secondary one.

Validation Images

Detection of Actin in tomato flower from wt plants and different RNAi lines (A) and olive root protein extracts from different cultivars (B) using ABIN3181106 diluted 1:2500 and 1:1000 respectively.

Protocole

Anticorps primaire: ABIN3181106

Anticorps secondaire: HRP conjugated rabbit anti-mouse IgG (Thermo Scientific, PA1-28902, OK1782904)

Full Protocol:

• Lyse protein extraction from 500mg olive root tissue and 80mg from tomato flower tissue in 240µl cold lysis buffer (1M Tris-Cl pH6.8, 10% SDS, 14.3M β-Merc, 0.5N EDTA, PMSF 0.5mM and 100µl proteinase inhibitor (Sigma-Aldrich, P9599-1ML)).
• Determine total protein content of the lysates using protein assay (Bradford curve).
• Denature 0.7μg/μl of total protein for 5min at 95°C 1:4 (buffer:protein) in Laemmli SDS sample buffer (Tris-Cl pH6.8, 2% SDS, 10% glycerol, 1%β-mercaptoethanol, 12.5mM EDTA, 0.02% bromophenol blue) and subsequently separate them on a freshly cast denaturing SDS-PAGE gel (30% Bis-Acrylamide, 1.5M Tris pH8.8, 10% SDS, 10% APS and TEMED) in an Bio-Rad Mini-PROTEAN 3 cell systemfor 2h at 100V, 200mA.
• Transfer proteins onto an Immobilon-P membrane (Millipore, IPVPH0010, K5CA2260G) with a Western blotting system for 1h at 100V.
• Block the membrane with 5% non-fat milk in 1x TBST for 1h at RT.
• Incubate membrane with anti-Actin antibody (antibodies-online, ABIN3181106, lot B3401) diluted 1:2500 (Flower samples) or 1:1000 (root samples) in 5% non-fat milk in 1x TBST ON at 4°C.
• Wash membrane 3x for 5min with 1x TBST.
• Incubate membrane with secondary HRP conjugated rabbit anti-mouse IgG (Thermo Scientific, PA1-28902, OK1782904) diluted 1: 5000 (flower samples) and 1:2000 (root samples) in 5% non-fat milk in 1x TBSTfor 1h at RT.
• Wash membrane 3x for 5min with 1X TBST.
• Reveal protein bands using super signal west pico (Thermo-Scientific, 34080, QF218483) on a Super RX Fuji Medical X-RAY (18cm x 24cm) film.

Notes:

The actin antibody ABIN3181106 reveals the actin protein of the expected molecular weight of antigen in lysates of tomato flower and olive root samples.

Stockage

Format: Liquid

Concentration: 1 mg/mL

Buffer: Liquid in PBS containing 50 % glycerol, 0.5 % BSA and 0.02 % sodium azide.

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Conseil sur la manipulation: Avoid repeated freeze/thaw cycles.

Stock: -20 °C

Stockage commentaire: Store at -20°C.

Références pour anticorps anti-Actin (ABIN3181106)

Hoffmann, Dietrich, Herrmann, Schacht, Albrecht, Methner: "Dimethyl Fumarate Induces Glutathione Recycling by Upregulation of Glutathione Reductase." dans: Oxidative medicine and cellular longevity, Vol. 2017, pp. 6093903, (2017) (PubMed).

Détails sur Actin

Antigène: Actin

Abstract: Actin Produits

Synonymes: anticorps Act57, anticorps Act57A, anticorps Act57b, anticorps Actin, anticorps BEST:RE38067, anticorps C, anticorps CG10067, anticorps CG30294, anticorps Dmel\\CG10067, anticorps M, anticorps T10, anticorps act 57A, anticorps act57A, anticorps act57B, anticorps act57b, anticorps actin, anticorps anon-EST:CL2c12, anticorps 143060_f_at, anticorps CG7478, anticorps D, anticorps Dmel\\CG7478, anticorps act 79B, anticorps act79B, anticorps mus79B, anticorps Act(88F), anticorps Act88-F, anticorps Act88f, anticorps CG5178, anticorps Dmel\\CG5178, anticorps E, anticorps Ifm(3)7, anticorps act 88F, anticorps act88F, anticorps rsd, anticorps Act87e, anticorps CG18290, anticorps Dmel\\CG18290, anticorps act 87E, anticorps act87E, anticorps mus87E, anticorps Act88F, anticorps Dsim\\GD19025, anticorps GD19025, anticorps dsim_GLEANR_2829, anticorps 42A, anticorps A, anticorps AFFX-Dros-ACTIN_M_r_at, anticorps Act, anticorps Act42, anticorps Act42a, anticorps Actin/BAP47, anticorps BAP47, anticorps CG12051, anticorps Dmel\\CG12051, anticorps act 42A, anticorps act42A, anticorps cytoplasmic, anticorps Actin 57B, anticorps Actin 79B, anticorps Actin 88F, anticorps Actin 87E, anticorps Actin-like protein C08B11.6, anticorps hypothetical protein, anticorps ACT_COLSC Actin (ISS), anticorps Actin 42A, anticorps actin gamma 1, anticorps actin, anticorps Actin, gamma 1, anticorps si:ch211-241j12.3, anticorps Act57B, anticorps Act79B, anticorps Act88F, anticorps Act87E, anticorps arp-6, anticorps Dsim\Act88F, anticorps ACTIN, anticorps Act42A, anticorps ACTG1, anticorps LOC548170, anticorps Bm1_16810, anticorps IDA5, anticorps LOC100342017, anticorps si:ch211-241j12.3