beta Actin Antikörper

Produktnummer ABIN387703

Produktdetails anti-beta Actin Antikörper

Target: beta Actin (ACTB) (Actin, beta (ACTB))

Reaktivität: Human, Maus, Huhn, Kaninchen, Schwein, Rind (Kuh), Hamster, Ziege, Fisch

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser beta Actin Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA

Spezifität: THE^TM beta Actin Antibody, mAb, Mouse reacts with mouse, rabbit, chicken, human, hamster, cow, goat, fish, and pig. It has not yet been tested in other species.

Kreuzreaktivität (Details): Cross reactivity in other species has not yet been tested.

Aufreinigung: Protein G affinity column

Immunogen: A synthetic peptide DDDIAALVVDNGSG coupled - KLH

Klon: 2D1D10

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: Working concentrations for specific applications should be determined by the investigator. The appropriate concentrations may be affected by secondary antibody affinity, antigen concentration, the sensitivity of the method of detection, temperature, the length of the incubations, and other factors. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product.
ELISA: 0.1-1.0 µg/mL
Western blot: 1.0 µg/mL
Other applications: user-optimized

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #102832 (Western Blotting)

by: Dr. Randy Brutkiewicz Laboratory, Department of Microbiology and Immunology, Indiana University School of Medicine

No.: #102832

Datum: 20.02.2018

Antigen: ACTB

Validierte Anwendung: Western Blotting

Positivkontrolle: lysates from HEK293 cells untransfected or transfected with human MR1 cDNA

Bewertung:

Passed. ABIN962807 recognizes human beta actin in HEK293 cell lysates.

Validierungsbilder

Lysates from human MR1-expressing HEK293 cells (MR1) and vector control cells (Vc) were resolved on a 10% SDS-PAGE gel for Western blotting analysis using antibodies specific for MR1 (ABIN1537116) and beta-actin (ABIN962807).

Protokoll

Primärantikörper: ABIN962807

Sekundärantikörper: goat anti-rabbit Dye-IR800 conjugated antibody (Advansta, R-05060-250, lot 17083179)

Full Protocol:

• Grow HEK293 cells in DMEM medium (Lonza, 12-614F, lot 0000618582) supplemented with serum (Hyclone, SH30071.03, lot AAG205460) and antibiotics (Hyclone, SV30010, lot J150013), at 37°C and 5% CO₂ dish to 70-90% confluency.
• Transfect cells with pCDNA 3.1 neo (-) (Invitrogen) containing human MR1 cDNA (Genecopoeia) using Polyethylenimine (Polysciences, 23966) following the manufacturer´s instructions.
• Lyse cells in cold lysis buffer (10mM Tris pH7.4, 150mM NaCl, 0.5mM EDTA, 2% CHAPS).
• Determine total protein content of the lysates using Commassie Protein Assay Reagent (Thermo Scientific, 1856209, lot NL179252).
• Denature 200µg total protein for 5min at 95°C in 20µl Laemmli SDS sample buffer and subsequently separate them on a SDS-PAGE gel using Acrylamide/Bis Premixed (Bio-Rad, 61-0125, lot 260000477) for 2-3h at 100V.
• Transfer proteins onto PVDF membrane (Millipore, IPVH00010, lot K5AA6843U) with a Western blotting system for ON at 4°C at 150mA.
• Block the membrane with blocking buffer (2% BSA/PBS/0.05%Tween-20) for 1h at RT.
• Incubate membrane with:
• loading control rabbit anti beta-actin (antibodies-online, ABIN962807) diluted 1:500 in blocking buffer ON at 4°C.
  • primary rabbit anti-MR1 antibody (antibodies-online, ABIN1537116, lot SA111213CH) diluted 1:1000 in blocking buffer ON at 4°C.
• Wash membrane 3x for 10min with PBS/0.05%Tween-20.
• Incubate membrane with secondary goat anti-rabbit Dye-IR800 conjugated antibody (Advansta, R-05060-250, lot 17083179) diluted 1:10000 in PBS/0.05% Tween-20 for 1h at RT.
• Wash membrane 3x for 10min with PBS/0.05% Tween-20.
• Reveal protein bands using an Odyssey imaging system (LI-COR Biosciences).

Anmerkungen:

The beta Actin antibody ABIN962807 reveals a protein of the expected molecular weight of beta actin in lysates of human HEK293 cells.

Handhabung

Format: Lyophilized

Rekonstitution: Reconstitute the lyophilized antibody with deionized water to the final concentration of 0.5 mg/mL.

Buffer: PBS, pH 7.4, containing 0.02 % Sodium azide, without BSA

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: The antibody is stable in lyophilized form if stored at -20°C or below. The reconstituted antibody can be stored for 2-3 weeks at 2-8°C. For long term storage, aliquot and store at -20°C or below. Avoid repeated freezing and thawing cycles.

Referenzen für anti-beta Actin Antikörper (ABIN387703)

Zhao, Mo, Li, Zou, Cheng, Sun, Xiong, Guan, Lei: "NOTCH-induced aldehyde dehydrogenase 1A1 deacetylation promotes breast cancer stem cells." in: The Journal of clinical investigation, (2014) (PubMed).

Srivastava, Reinoso, Singh, Fernandez-Bueno, Martino, Garcia-Gutierrez, Pastor, Corell: "Flow cytometry assessment of the purity of human retinal pigment epithelial primary cell cultures." in: Journal of immunological methods, Vol. 389, Issue 1-2, pp. 61-8, (2013) (PubMed).

Details zu beta Actin

Target: beta Actin (ACTB) (Actin, beta (ACTB))

Andere Bezeichnung: beta Actin (ACTB Produkte)

Synonyme: BRWS1 antikoerper, PS1TP5BP1 antikoerper, ACTA1 antikoerper, Actx antikoerper, E430023M04Rik antikoerper, beta-actin antikoerper, ps1tp5bp1 antikoerper, act-2 antikoerper, actb antikoerper, B-actin antikoerper, LOC100174883 antikoerper, LOC100304816 antikoerper, A antikoerper, ACT5C antikoerper, Ac5C antikoerper, Act antikoerper, Act-5C antikoerper, Act5 antikoerper, Act5c antikoerper, Actin antikoerper, Actin/BAP47 antikoerper, Actin5C antikoerper, BAP47 antikoerper, Bap47 antikoerper, CG4027 antikoerper, Dmel\\CG4027 antikoerper, M32055 antikoerper, T11 antikoerper, act antikoerper, act 5C antikoerper, act5C antikoerper, actin antikoerper, actin5C antikoerper, anon-EST:fe2D2 antikoerper, beta-actin/Bap47 antikoerper, cyt5C antikoerper, l(1)G0009 antikoerper, l(1)G0010 antikoerper, l(1)G0025 antikoerper, l(1)G0079 antikoerper, l(1)G0117 antikoerper, l(1)G0177 antikoerper, l(1)G0245 antikoerper, l(1)G0330 antikoerper, l(1)G0420 antikoerper, l(1)G0486 antikoerper, Bact antikoerper, ACT-5 antikoerper, ACTB antikoerper, B-ACTZF antikoerper, actba antikoerper, bact antikoerper, bactin1 antikoerper, bactzf antikoerper, wu:fd18f05 antikoerper, A26C1B antikoerper, POTE2alpha antikoerper, POTEACTIN antikoerper, Beta-actin antikoerper, actin beta antikoerper, actin, beta antikoerper, cytoplasmic actin OlCA1 antikoerper, beta-actin antikoerper, Actin 5C antikoerper, actin gamma 1 antikoerper, actin, beta 1 antikoerper, actin, beta L homeolog antikoerper, beta actin antikoerper, POTE ankyrin domain family member F antikoerper, actin, beta like 2 antikoerper, actin, cytoplasmic 1 antikoerper, ACTB antikoerper, Actb antikoerper, actb antikoerper, LOC100174883 antikoerper, LOC100304816 antikoerper, Act5C antikoerper, ACTG1 antikoerper, actb1 antikoerper, actb.L antikoerper, LOC100136352 antikoerper, POTEF antikoerper, ACTBL2 antikoerper, LOC109073280 antikoerper

Hintergrund: Beta-actin is one of the six different actin isoforms that have been identified. The actin molecules found in cells of various species and tissues tend to be very similar in their immunological and physical properties. As a consequence, it has been difficult to produce potent antisera against this protein. Therefore, the availability of monoclonal antibodies to beta-actin provides a useful and specific tool in the study of the intracellular distribution of beta-actin and the static and dynamic aspects of the cytoskeleton.

Pathways: Myometrial Relaxation and Contraction, Cell-Cell Junction Organization, Maintenance of Protein Location, Phototransduction