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beta Actin 抗体 (AA 2-16)

Name: beta Actin 抗体 (AA 2-16)
SKU: ABIN1742508
Availability: OutOfStock
Rating: 5 (1 reviews)

ACTB 适用: 小鼠, 大鼠, 小鸡, 斑马鱼 WB, IHC, ICC 宿主: 兔 Polyclonal unconjugated

Western Blotting validation image for anti-Actin, beta (ACTB) (AA 2-16) antibody (ABIN1742508) (beta Actin 抗体 (AA 2-16))

Western Blotting (WB) image for anti-Actin, beta (ACTB) (AA 2-16) antibody (ABIN1742508) (beta Actin 抗体 (AA 2-16))

Immunocytochemistry (ICC) image for anti-Actin, beta (ACTB) (AA 2-16) antibody (ABIN1742508) (beta Actin 抗体 (AA 2-16))

ICC

Immunohistochemistry (IHC) image for anti-Actin, beta (ACTB) (AA 2-16) antibody (ABIN1742508) (beta Actin 抗体 (AA 2-16))

IHC

Indirect immunofluorescence labeling of PFA fixed, paraffin embedded mouse muscle section (dilution 1 : 500; red). (beta Actin 抗体 (AA 2-16))

IHC (p)

7 images

(3 references)

(1 validation)

产品编号 ABIN1742508

发货至: 中国

Datasheet as PDF

抗原 See all beta Actin (ACTB) 抗体

beta Actin (ACTB) (Actin, beta (ACTB))
抗原表位
35

17

16

6

6

5

5

3

3

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1
AA 2-16
适用
180

129

117

63

49

44

37

36

32

24

20

20

12

10

9

8

7

6

5

5

3

3

3

3

2

2

2

2

1

1

1

1

1

1

1

1

1
小鼠, 大鼠, 小鸡, 斑马鱼
宿主
106

89

5

2

1
兔
Compatible Secondaries
克隆类型
106

95
多克隆
标记
124

14

14

9

4

4

4

4

4

4

3

2

2

2

1

1

1

1

1

1

1

1

1
This beta Actin antibody is un-conjugated
应用范围
170

71

54

49

33

29

25

21

13

13

7

6

6

5

3

2

1

1
Western Blotting (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC)

交叉反应（详细）

May cross-react to alpha- and gamma-actin due to sequence homology.

纯化方法

Affinity purified with the immunogen. Rabbit serum albumin was added for stabilization.

免疫原

Synthetic peptide (aa 2-16 of mouse beta-actin) coupled to KLH via a C-terminally added cysteine.

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anti-Actin, beta (ACTB) (AA 1-50) antibody
ACTB 适用: 人 WB, ELISA, IHC, IF, IP, FACS 宿主: 小鼠 Monoclonal 1D12E12B5 unconjugated

应用备注

WB: 1 : 1000 up to 1 : 5000
IP: not tested yet
ICC: 1 : 200 up to 1 : 500
IHC: 1 : 200

说明

ICC: Methanol fixation is recommended.

限制

仅限研究用
生效 #029836 (Western Blotting)

by

Kinexus Bioinformatics Corporation

No.

#029836

日期

2015.06.22

抗原

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)

Validation Images

Figure 1. Western blot for beta actin (ACTB). Grey arrowhead indicates the expected molecular weight of ~42 kDa.

Figure 2. Ponceau stain loading control.

Figure 3. Western blot for beta actin (ACTB) using attempted siRNA silencing. Arrowhead indicates the expected molecular weight of ~42 kDa. HeLa cells were transfected with beta actin siRNA (Catalog number 12935141, Life Technologies) using Lipofectamine 2000, according to manufacturer's instructions. Total cell lysates were made from both floating and adherent cells harvested 48 hours and 72 hours after transfection. siRNA silencing of beta actin was not detected.
Full Methods
Primary Antibody

Antigen: Beta-Actin, (ACTB) (AA 2-16)
Catalog number: ABIN1742553
Lot number: 251003/3
Dilution: 1:1000

Secondary Antibody

Antibody: Donkey anti-Rabbit IgG Antibody (HRP)
Lot number: F0613
Dilution: 1:10,000

Full 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.
状态

Lyophilized

溶解方式

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.

缓冲液

PBS

注意事项

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.

储存条件

-20 °C

储存方法

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!
Lu, Ma, Zhuang, Zhang, Cheng, Shi: "Retinoic acid‑incorporated glycol chitosan nanoparticles inhibit the expression of Ezh2 in U118 and U138 human glioma cells." in: Molecular medicine reports, Vol. 12, Issue 5, pp. 6642-8, (2016) (PubMed).

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).
抗原

beta Actin (ACTB) (Actin, beta (ACTB))

别名

beta-Actin (ACTB 产品)

别名

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

途径

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