beta Actin antibody (Actin, beta)

Details for Product anti-ACTB Antibody No. ABIN962807, Supplier: Log in to see
Antigen
  • BRWS1
  • PS1TP5BP1
  • ACTA1
  • Actx
  • E430023M04Rik
  • beta-actin
  • ps1tp5bp1
  • act-2
  • actb
  • B-actin
  • LOC100174883
  • LOC100304816
  • A
  • ACT5C
  • Ac5C
  • Act
  • Act-5C
  • Act5
  • Act5c
  • Actin
  • Actin/BAP47
  • Actin5C
  • BAP47
  • Bap47
  • CG4027
  • Dmel\\CG4027
  • M32055
  • T11
  • act
  • act 5C
  • act5C
  • actin
  • actin5C
  • anon-EST:fe2D2
  • beta-actin/Bap47
  • cyt5C
  • l(1)G0009
  • l(1)G0010
  • l(1)G0025
  • l(1)G0079
  • l(1)G0117
  • l(1)G0177
  • l(1)G0245
  • l(1)G0330
  • l(1)G0420
  • l(1)G0486
  • Bact
  • ACT-5
  • ACTB
  • B-ACTZF
  • actba
  • bact
  • bactin1
  • bactzf
  • wu:fd18f05
  • Beta-actin
  • actin beta
  • actin, beta
  • cytoplasmic actin OlCA1
  • beta-actin
  • Actin 5C
  • actin gamma 1
  • actin, beta 1
  • actin, beta L homeolog
  • beta actin
  • actin, cytoplasmic 1
  • ACTB
  • Actb
  • actb
  • LOC100174883
  • LOC100304816
  • Act5C
  • ACTG1
  • actb1
  • actb.L
  • LOC100136352
  • LOC109073280
Alternatives
anti-Cow (Bovine) beta Actin antibody for Immunocytochemistry
Reactivity
Pig (Porcine), Fish, Goat, Cow (Bovine), Hamster, Rabbit, Mouse (Murine), Human, Chicken
593
347
305
108
85
83
63
57
54
45
42
41
34
32
28
22
15
15
15
15
10
9
8
7
7
6
5
4
3
3
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
Host
Mouse
359
259
12
7
2
1
Clonality (Clone)
Monoclonal ()
Conjugate
This beta Actin antibody is un-conjugated
34
29
19
5
5
4
4
4
4
4
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
Application
ELISA, Western Blotting (WB)
579
251
183
134
115
97
60
45
37
15
14
13
12
9
9
2
2
2
2
2
1
1
1
1
Options
Supplier
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Supplier Product No.
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'Independent Validation' Badge
Antigen ACTB
Method validated Western Blotting
Positive Control lysates from HEK293 cells untransfected or transfected with human MR1 cDNA
Notes

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

Primary Antibody ABIN962807
Secondary Antibody goat anti-rabbit Dye-IR800 conjugated antibody (Advansta, R-05060-250, lot 17083179)
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).
Experimental Notes

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

Validation Images
Western Blotting validation image for anti-Actin, beta (ACTB) antibody (ABIN962807) Lysates from human MR1-expressing HEK293 cells (MR1) and vector control cells (Vc) we...
Immunogen A synthetic peptide DDDIAALVVDNGSG coupled - KLH
Clone 2D1D10
Isotype IgG
Specificity THETM 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.
Cross-Reactivity (Details) Cross reactivity in other species has not yet been tested.
Purification Protein G affinity column
Plasmids, Primers & others Plasmids, Primers & others beta Actin products on genomics-online (e.g. as negative or positive controls)
Antigen
Alternative Name beta Actin (ACTB Antibody Abstract)
Background 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
Application Notes 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
Restrictions For Research Use only
Format Lyophilized
Reconstitution 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
Preservative Sodium azide
Precaution of Use 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.
Storage 4 °C/-20 °C
Storage Comment 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.
Supplier Images
Western Blotting (WB) image for anti-Actin, beta (ACTB) antibody (ABIN962807) Lane 1: 20 µg Hela cell lysate Lane 2: 20 µg sp2/0 cell lysate Lane 3: 20 µg goat mus...
Western Blotting (WB) image for anti-Actin, beta (ACTB) antibody (ABIN962807) Primary antibody: A. 0.1 µg/mL Anti-beta-actin Monoclonal Antibody (Mouse) (ABIN39685...
Western Blotting (WB) image for anti-Actin, beta (ACTB) antibody (ABIN962807) Primary antibody: A. 0.1 µg/mL Anti-beta-actin Monoclonal Antibody (Mouse) (ABIN39685...
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