Anti-beta Actin 抗体 [mAbcam 8224] - Loading Control (ab8224)
Key features and details
- Mouse monoclonal [mAbcam 8224] to beta Actin - Loading Control
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human, Xenopus laevis, Drosophila melanogaster, Schizosaccharomyces pombe
- Isotype: IgG1
Related conjugates and formulations
製品の概要
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製品名
Anti-beta Actin antibody [mAbcam 8224] - Loading Control
beta Actin 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [mAbcam 8224] to beta Actin - Loading Control -
由来種
Mouse -
特異性
Recognises a single band at 42kD representing beta 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.
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アプリケーション
適用あり: WB, IHC-P, ICC/IFmore details
適用なし: Flow Cyt (Intra) -
種交差性
交差種: Mouse, Rat, Human, Xenopus laevis, Drosophila melanogaster, Schizosaccharomyces pombe
交差が予測される動物種: Rabbit, Chicken, Cow, Cat, Dog, Pig, Chinese hamster, Other species -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab13772) -
ポジティブ・コントロール
- WB: A431; HEK293; NIH3T3; PC12 whole cell lysates; Xenopus embryo lysate; Drosophila lysate; S. pombe lysate. ICC/IF: HeLa cells. IHC/P: Human colon (FFPE)
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特記事項
This monoclonal antibody to beta actin works well as a protein loading control in Western blot for a broad range of species including Xenopus, Drosophila and S. pombe.
This antibody clone [mAbcam 8224] 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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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精製度
Protein G purified -
一次抗体 備考
This clone works well as a loading control for Xenopus, Drosophila, S. cerevisiae and S.pombe. We recommend using ab8224 instead of ab8226 for these species. -
ポリ/モノ
モノクローナル -
クローン名
mAbcam 8224 -
ミエローマ
Sp2/0-Ag14 -
アイソタイプ
IgG1 -
軽鎖の種類
kappa -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab8224の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB | (22) |
Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin peptide (ab13772). This antibody has been designed for use as a loading control and is ideal for this purpose. Block membrane for 1 hr in 5%BSA. Incubate antibody in TBST for one hour or more.
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IHC-P | (7) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (6) |
Use a concentration of 1 µg/ml.
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特記事項 |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin peptide (ab13772). This antibody has been designed for use as a loading control and is ideal for this purpose. Block membrane for 1 hr in 5%BSA. Incubate antibody in TBST for one hour or more. |
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 1 µg/ml. |
ターゲット情報
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機能
Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. -
関連疾患
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. -
配列類似性
Belongs to the actin family. -
翻訳後修飾
ISGylated. -
細胞内局在
Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. - Information by UniProt
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参照データベース
- Entrez Gene: 396526 Chicken
- Entrez Gene: 280979 Cow
- Entrez Gene: 403580 Dog
- Entrez Gene: 60 Human
- Entrez Gene: 11461 Mouse
- Entrez Gene: 414396 Pig
- Entrez Gene: 100009272 Rabbit
- Entrez Gene: 81822 Rat
see all -
別名
- A26C1A antibody
- A26C1B antibody
- ACTB antibody
see all
画像
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All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1 µg/ml
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
Lane 5 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue
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 ab8224 overnight at 4°C. Antibody binding was detected using a goat anti-mouse 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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IHC image of ab8224 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 ab8224, 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 -
ab8224 staining beta Actin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab8224 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 3 minutesThis 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 2% Bovine Serum Albumin before being incubated with ab8224 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406
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IHC image of beta actin staining in human colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8224, 3µg/ml overnight at +4°C. A goat anti-mouse HRP-conjugated secondary antibody (ab6789, 1/2000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
The inset negative control image is secondary-only at 1/500 dilution.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) + Xenopus embryo lysate at 20 µg
Secondary
Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDaab8224 used on Xenopus embryo lysate (20 ug of lysate/lane).
Secondary
Rabbit polyclonal anti-mouse HRP was used as the secondary antibody (ab6728) and developed using the ECL technique.
Performed under reducing conditions.
Predicted band size : 42kD -
All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1 µg/ml
Lane 1 : Drosophila lysate
Lane 2 : S. pombe lysate
Lane 3 : S. cerevisiae lysate (Actin 1 - please see note)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDaNote: although S. cerevisiae is not known to express beta Actin, Abcam believes that the band on lane 3 corresponds to Actin 1 (Swissprot ID: P60010, based on sequence similarity).
Secondary antibody - rabbit anti-mouse HRP (ab6728)
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All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1/1000 dilution
Lane 1 : Fruit fly (Drosophila melanogastor) whole cell lysate - Female
Lane 2 : Fruit fly (Drosophila melanogastor) whole cell lysate -
Male
Lysates/proteins at 100 µg per lane.
Secondary
All lanes : An HRP-conjugated Sheep polyclonal to mouse IgG at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Exposure time: 2 minutes
Blocking step: 5% Milk for 1 hour at 20°C. -
Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1/10000 dilution + Mouse CT26 cells at 40 µg
Secondary
Anti-mouse IgG, HRP-linked Antibody at 1/10000 dilution
Developed using the ECL technique.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 6 minutesWestern blot analysis using ab8224 at 1:1000 on Mouse CT26 cells. Blocking agent and dilution buffer was 5% milk in TBST
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Immunohistochemistical detection of beta Actin using antibody [mAbcam 8224] - Loading Control on formaldehyde-fixed paraffin-embedded rat cerebellum sections. Antigen retrieval step: heat mediated Citric acid pH6 buffer. Permeabilization: No. Blocking step: 1% BSA for 10 mins @ rt°C. Primary antibody dilution 1/1000 for 2 hours in TBS/BSA/azide. Secondary Antibody: anti Mouse Igs conjugated to biotin (1/200). beta Actin appears to be particularly enriched not only in the glomeruli of the Granule cell layer (indicated by red arrowheads ) but also in Microglia (indicated by green arrowheads); All positive microglia appear to be ramified thus not presumed to be activated.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (472)
ab8224 は 472 報の論文で使用されています。
- Katz B et al. Nociception and pain in humans lacking a functional TRPV1 channel. J Clin Invest 133:N/A (2023). PubMed: 36454632
- Golchoubian B et al. Reticulon-like REEP4 at the inner nuclear membrane promotes nuclear pore complex formation. J Cell Biol 221:N/A (2022). PubMed: 34874453
- Drazic A et al. The Final Maturation State of β-actin Involves N-terminal Acetylation by NAA80, not N-terminal Arginylation by ATE1. J Mol Biol 434:167397 (2022). PubMed: 34896361
- Nguyen N et al. Recruitment of MLL1 complex is essential for SETBP1 to induce myeloid transformation. iScience 25:103679 (2022). PubMed: 35036869
- Haraguchi T et al. Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase. Commun Biol 5:78 (2022). PubMed: 35058555