beta Actin antibody (Actin, beta) (AA 2-16)

Details for Product anti-ACTB Antibody No. ABIN1742508, Supplier: Log in to see
Antigen
  • beta-actin
  • ps1tp5bp1
  • LOC563571
  • ACTB
  • A26C1A
  • A26C1B
  • act-2
  • actb
  • B-actin
  • LOC100136013
  • 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
  • BRWS1
  • PS1TP5BP1
  • ACT-5
  • Actx
  • E430023M04Rik
  • ACTA1
  • Beta-actin
  • B-ACTZF
  • actba
  • bact
  • bactin1
  • bactzf
  • wu:fd18f05
  • POTE2alpha
  • POTEACTIN
  • actin, beta
  • similar to beta-cytoplasmic actin2
  • cytoplasmic actin OlCA1
  • actin beta
  • beta-actin
  • Actin 5C
  • actin, beta 1
  • actin, gamma 1
  • beta actin
  • POTE ankyrin domain family, member F
  • actb
  • LOC563571
  • ACTB
  • LOC100136013
  • LOC100174883
  • LOC100304816
  • Act5C
  • Actb
  • actb1
  • ACTG1
  • LOC396797
  • LOC100350495
  • LOC100136352
  • POTEF
Alternatives
anti-Mouse (Murine) beta Actin antibody for Immunofluorescence (fixed cells)
Epitope
AA 2-16
163
40
17
17
11
9
9
4
4
3
3
3
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
Reactivity
Chicken, Mouse (Murine), Rat (Rattus), Zebrafish
571
346
298
103
82
74
59
58
45
42
42
37
32
28
22
16
15
13
13
13
9
9
8
7
7
6
5
4
4
3
3
2
2
2
2
1
1
1
1
1
1
1
1
1
1
Host
Rabbit
354
233
16
8
1
1
Clonality
Polyclonal
Conjugate
This beta Actin antibody is un-conjugated
32
27
18
6
6
5
5
4
4
4
3
3
3
3
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
Immunocytochemistry (ICC), Immunohistochemistry (IHC), Western Blotting (WB)
544
227
155
135
121
98
59
46
19
14
13
12
10
9
9
2
2
2
2
2
1
1
1
1
Options
Supplier
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Supplier Product No.
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Available images

'Independent Validation' Badge
Lot Number 251003/3
Method validated Western Blotting
Positive Control mouse brain, mouse kidney, mouse muscle, mouse spleen, mouse testes
Negative Control none [ubiquitously expressed]
Notes A band was observed in the positive controls at the expected size (~42 kDa)
Primary Antibody
  • Antigen: Beta-Actin, (ACTB) (AA 2-16)
  • Catalog number: ABIN1742553
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: 251003/3
  • Dilution: 1:1000
Secondary Antibody
  • Antibody: Donkey anti-Rabbit IgG Antibody (HRP)
  • Catalogue number: Log in to see
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  • Lot number: F0613
  • Dilution: 1:10,000
Controls
  • All lysates used were prepared by Kinexus Bioinformatics Corporation following standard protocols and quality controlled for protein integrity on a regular basis.
  • Loading control: blot was stripped and re-probed with β3-Tubulin.
Protocol
  • 1. Cell/tissue total protein lysates were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 minutes prior to loading.
  • 2. 15 μg of boiled lysate were loaded and resolved on a 12% SDS-polyacrylamide gel.
  • 3. The Precision Plus Protein™ All Blue Prestained Standards from BioRad (161-0373) were used as molecular mass markers.
  • 4. Proteins were transferred onto nitrocellulose membrane by tank transfer and protein transfer was confirmed with Ponceau S staining.
  • 5. The immunoblot membrane was blocked in 2.5% skim milk and 1.5% BSA solution in TTBS at room temperature for 60 minutes.
  • 6. The membrane was washed in TTBS twice for 5 minutes each.
  • 7. The membrane was immersed with the protein side up in the antibody solution in TBS and incubated overnight at 4°C with gentle agitation.
  • 8. The membrane was rinsed twice with TTBS.
  • 9. The membrane was washed in TTBS twice for 5 minutes each.
  • 10. The membrane was washed in TTBS once for 15 minutes.
  • 11. The membrane was incubated in the HRP-conjugated secondary antibody solution in TBS for 60 minutes at room temperature with gentle agitation.
  • 12. The membrane was rinsed twice with TTBS.
  • 13. The membrane was washed in TTBS twice for 5 minutes each.
  • 14. The membrane was washed in TTBS once for 15 minutes.
  • 15. Signals were detected by chemiluminescence (ECL). The blot was scanned for 160 seconds.
  • 16. The membrane was rinsed three times with TTBS.
  • 17. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 minutes each.
  • 18. The membrane was washed in TTBS 3 times for 5 minutes each.
  • 19. Repeated Steps 4-14 with the loading control antibody (for β3−Tubulin).
  • 20. Repeated Steps 4-14 with the loading control antibody and its matching secondary antibody. The blot was scanned for 160 seconds.
Experimental Notes A protein band was observed at the expected target size of β−Actin of 42 kDa in all mouse extracts tested. Lower molecular weight bands of approximately 28-30 kDa were also observed in the mouse kidney and spleen tissue samples. Three main groups of actin have been identified. The β-actins and γ-actins co-exist in most cell types as components of the cytoskeleton, whereas the α-actin are found in muscle tissues. In our experience, we find that the level of actin does vary widely between different cell and tissue types.
Validation Images
Western Blotting validation image for anti-Actin, beta (ACTB) (AA 2-16) antibody (ABIN1742508) Figure 1. Western blot for beta actin (ACTB). Grey arrowhead indicates the expected m...
Western Blotting validation image for anti-Actin, beta (ACTB) (AA 2-16) antibody (ABIN1742508) Figure 2. Ponceau stain loading control.
Western Blotting validation image for anti-Actin, beta (ACTB) (AA 2-16) antibody (ABIN1742508) Figure 3. Western blot for beta actin (ACTB) using attempted siRNA silencing. Arrowhe...
Immunogen Synthetic peptide (aa 2-16 of mouse beta-actin) coupled to KLH via a C-terminally added cysteine.
Cross-Reactivity (Details) May cross-react to alpha- and gamma-actin due to sequence homology.
Purification Affinity purified with the immunogen. Rabbit serum albumin was added for stabilization.
Alternative Name beta-Actin (ACTB Antibody Abstract)
Pathways Myometrial Relaxation and Contraction, Cell-Cell Junction Organization, Maintenance of Protein Location, Phototransduction
Application Notes WB: 1 : 1000 up to 1 : 5000
IP: not tested yet
ICC: 1 : 200 up to 1 : 500
IHC: 1 : 200
Comment

ICC: Methanol fixation is recommended.

Restrictions For Research Use only
Format Lyophilized
Reconstitution For reconstitution add 100 µL H2O to get a 1mg/ml solution of antibody in PBS. Then aliquot and store at -20 °C until use.
Buffer PBS
Handling Advice Affinity purified antibodies are less robust than antisera, since protease inhibitors are also removed during purification. Hence, storage at 4 °C for prolonged periods (i.e. several weeks), is not recommended.
Storage -20 °C
Storage Comment Unlabeled lyophilized antibodies are stable in this form without loss of quality at ambient temperatures for several weeks or even months. They can be stored at 4°C for several years. Lyophilized antibodies must not be stored in the freezer, they may be destroyed!
Supplier Images
Western Blotting (WB) image for anti-beta Actin antibody (Actin, beta) (AA 2-16) (ABIN1742508) anti-Actin, beta (ACTB) (AA 2-16) antibody
Immunocytochemistry (ICC) image for anti-beta Actin antibody (Actin, beta) (AA 2-16) (ABIN1742508) anti-Actin, beta (ACTB) (AA 2-16) antibody (Image 2)
Immunohistochemistry (IHC) image for anti-beta Actin antibody (Actin, beta) (AA 2-16) (ABIN1742508) anti-Actin, beta (ACTB) (AA 2-16) antibody (Image 3)
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-beta Actin antibody (Actin, beta) (AA 2-16) (ABIN1742508) Indirect immunofluorescence labeling of PFA fixed, paraffin embedded mouse muscle sec...
Product cited in: Baron, Blex, Rohrbeck, Rachakonda, Birnbaumer, Ahnert-Hilger, Brunk: "The α-subunit of the trimeric GTPase Go2 regulates axonal growth." in: Journal of neurochemistry, Vol. 124, Issue 6, pp. 782-94, 2013 (PubMed).

Whitemarsh, Strathman, Chase, Stankewicz, Tepp, Johnson, Pellett: "Novel application of human neurons derived from induced pluripotent stem cells for highly sensitive botulinum neurotoxin detection." in: Toxicological sciences : an official journal of the Society of Toxicology, Vol. 126, Issue 2, pp. 426-35, 2012 (PubMed).