Anti-beta Actin抗体[mAbcam 8226] - Loading Control (ab8226)
- 数据表
- 参考文献 (1234)
- 实验方案
概述
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产品名称
Anti-beta Actin抗体[mAbcam 8226] - Loading Control
参阅全部 beta Actin 一抗 -
描述
小鼠单克隆抗体[mAbcam 8226] to beta Actin - Loading Control -
宿主
Mouse -
特异性
Does not cross-react with adult cardiac, smooth, or skeletal muscle actin. The immunogen used for this product shares 77% homology with Gamma actin/actin cytoplasmic 2. Cross-reactivity with this protein has not been confirmed experimentally. -
经测试应用
适用于: ICC/IF, ICC, Flow Cyt, IHC-FrFl, IHC-P, IHC-Fr, IP, WBmore details -
种属反应性
与反应: Mouse, Rat, Rabbit, Horse, Chicken, Cow, Dog, Human, Pig, Zebrafish, African green monkey, Chinese hamster, Armenian hamster
预测可用于: Sheep, Guinea pig -
免疫原
Synthetic peptide corresponding to Human beta Actin aa 1-100 conjugated to keyhole limpet haemocyanin (Sulfosuccinimidyl 4-N-maleimidomethyl-cyclohexane-1-carboxylate (Sulfo-SMCC)).
(Peptide available asab13772) -
阳性对照
- WB, ICC/IF, Flow Cyt: HeLa, Jurkat, A431, HEK293, NIH 3T3, PC12 cells. IHC-P: Human colon (FFPE), Rat colon (FFPE).
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常规说明
Western blot protocol advice:
We recommend blocking with 2-5% BSA as we have found that use of 5% milk significantly reduces the band intensity for beta actin. Please see the comparison data in the images section. If milk block is required, we recommend using ab8224 mouse monoclonal [mAbcam 8224] to beta actin. Contact our Scientific Support team for more information or advice.
This antibody clone [mAbcam 8226] is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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纯度
IgG fraction -
克隆
单克隆 -
克隆编号
mAbcam 8226 -
骨髓瘤
Sp2/0-Ag14 -
同种型
IgG1 -
轻链类型
kappa -
研究领域
相关产品
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Alternative Versions
- Anti-beta Actin antibody [mAbcam 8226] - Loading Control (Alexa Fluor® 680) (ab184092)
- Anti-beta Actin antibody [mAbcam 8226] - Loading Control (Alexa Fluor® 790) (ab184576)
- Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272)
- Anti-Actin antibody [mAbGEa] (ab230169)
- Anti-beta Actin antibody [mAbcam 8226] - BSA and Azide free (ab264083)
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Immunohistochemistry kits
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Isotype control
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Recombinant Protein
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Related Products
应用
Our Abpromise guarantee covers the use of ab8226 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
应用 | Ab评论 | 说明 |
---|---|---|
ICC/IF | Use a concentration of 5 µg/ml. | |
ICC | Use at an assay dependent concentration. | |
Flow Cyt | Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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IHC-FrFl | Use at an assay dependent concentration. | |
IHC-P | Use a concentration of 0.1 - 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
IHC-Fr | 1/500. | |
IP | 1/100. | |
WB | Use a concentration of 1 µg/ml. Predicted molecular weight: 42 kDa.Can be blocked with Human beta Actin peptide (ab13772). We recommend blocking using 2-5% BSA as we have found that use of 5% milk significantly reduces the band intensity for beta actin. Please refer to the images section for the blocking comparison data. |
靶标
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功能
Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. -
疾病相关
Defects in ACTB are a cause of dystonia juvenile-onset (DYTJ) [MIM:607371]. DYTJ is a form of dystonia with juvenile onset. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYTJ patients manifest progressive, generalized, dopa-unresponsive dystonia, developmental malformations and sensory hearing loss. -
序列相似性
Belongs to the actin family. -
翻译后修饰
ISGylated. -
细胞定位
Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. - Information by UniProt
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数据库链接
- Entrez Gene: 396526 Chicken
- Entrez Gene: 280979 Cow
- Entrez Gene: 403580 Dog
- Entrez Gene: 100033878 Horse
- Entrez Gene: 60 Human
- Entrez Gene: 11461 Mouse
- Entrez Gene: 414396 Pig
- Entrez Gene: 100009272 Rabbit
see all -
别名
- A26C1A antibody
- A26C1B antibody
- ACTB antibody
see all
图片
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All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at 1/10000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDaThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Immunofluorescence using ab8226 at 5µg/ml incubated for 1 hour on Rat Colon Cancer cells.
Cells were fixed with ice-cold methanol for 5 mins, then for all following steps, permeabilised in TBS-T for 30 mins, blocked with 5% BSA for 30 mins and then washed in TBS-T. Secondary antibody was Alexa Fluor 488 goat anti-mouse IgG at 1/1000 incubated for 1 hour. Cells were counterstained with DAPI. Image at 400X magnification. All incubations were at room temperature.
The beta actin fibres can be seen arrayed around the edge of the cells.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
IHC image of ab8226 staining beta Actin in human colon formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8226, 0.1μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
IHC image of ab8226 staining beta Actin in rat colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8226, 0.5μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Lane 1 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lane 2 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/10000 dilution
Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/500 dilution
Lanes 1-2 : HeLa cell lysate
Lane 3 : HEK293 cell lysate
Lane 4 : NIH 3T3 mouse cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Exposure time: 10 seconds -
All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 10 secondsWestern blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) with the Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623).
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Lanes 1-3 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% BSA BLOCK)
Lanes 4-6 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% MILK BLOCK)
Lanes 1 & 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lanes 2 & 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lanes 3 & 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 8 minutes -
Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 0.5 µg/ml + HeLa cell lysate
Secondary
Goat polyclonal to mouse IgG H&L (HRP) at 1/5000 dilution
Performed under non-reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 30 seconds -
Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)This image is a courtesy of Anonymous Abreview
ab8226 staining beta Actin in human gastric epithelial AGS cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde. Permeabilization and blocking was carried out using 5% BSA containing 0.025% Triton X in TBS for 1 hour at 23°C. Primary antibody was used at 5µg/ml for 1 hour at 23°C. An Alexa Fluor 488® conjugated goat polyclonal to mouse IgG was used as the secondary antibody at a 1/1000 dilution.
文献
This product has been referenced in:
- Yu J et al. DNA hypomethylation promotes invasion and metastasis of gastric cancer cells by regulating the binding of SP1 to the CDCA3 promoter. J Cell Biochem 121:142-151 (2020). Read more (PubMed: 31211445) »
- Zhang J et al. Upregulation of LSD1 promotes migration and invasion in gastric cancer through facilitating EMT. Cancer Manag Res 11:4481-4491 (2019). Read more (PubMed: 31191010) »