Actin Antikörper

Produktnummer ABIN3181106

Produktdetails anti-Actin Antikörper

Target: Actin

Reaktivität: Pflanzen

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser Actin Antikörper ist unkonjugiert

Applikation: Western Blotting (WB)

Spezifität: The antibody detects endogenous plant actin protein.

Produktmerkmale: Mouse Monoclonal to Plant actin.

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

Klon: Q30

Isotyp: IgG1

Anwendungsinformationen

Applikationshinweise: WB 1:2000-5000

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #102343 (Western Blotting)

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

No.: #102343

Datum: 19.12.2017

Antigen: Actin

Chargennummer: B3401

Validierte Anwendung: Western Blotting

Positivkontrolle: protein extraction from olive root tissue and tomato flower tissue

Bewertung:

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.

Validierungsbilder

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.

Protokoll

Primärantikörper: ABIN3181106

Sekundärantikörper: 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.

Anmerkungen:

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

Handhabung

Format: Liquid

Konzentration: 1 mg/mL

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

Konservierungsmittel: Sodium azide

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

Handhabung: Avoid repeated freeze/thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20°C.

Referenzen für anti-Actin Antikörper (ABIN3181106)

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

Details zu Actin

Target: Actin

Abstract: Actin Produkte

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