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.
Mouse, Rat, Human
预测可用于: Sheep, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Pig, Monkey, Zebrafish, African green monkey, Chinese hamster, Armenian hamster
Synthetic peptide corresponding to Human beta Actin aa 1-100 conjugated to keyhole limpet haemocyanin (Sulfosuccinimidyl 4-N-maleimidomethyl-cyclohexane-1-carboxylate (Sulfo-SMCC)).
Database link: P60709
(Peptide available as ab13772)
WB, ICC/IF: HeLa, Jurkat, A431, HEK293, NIH 3T3, PC12 cells.IHC-P: Human colon (FFPE), Rat colon (FFPE) tissue. Hela cells
Western blot protocol advice:
Werecommend blocking with 2-5% BSA as we have found that use of 5% milk significantly reduces theband intensityfor beta actin. Please see the comparison data in the images section. If milk block is required, we recommend usingab8224 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 aparticular buffer formulation or aparticular conjugate for your experiments, pleasecontactorders@abcam.comor you can findfurther informationhere.
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
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
The Abpromise guarantee
Abpromise™承诺保证使用ab8226于以下的经测试应用
\"应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
IHC-P (1) 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.
Use a concentration of 1 µg/ml. Predicted molecular weight: 42 kDa.Can be blocked with Human beta Actin peptide (ab13772).
Werecommend blocking using 2-5% BSA as we have found that use of 5% milk significantly reduces theband intensityfor beta actin. Please refer to the images section for the blocking comparison data.
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.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 42 kDa.Can be blocked with Human beta Actin peptide (ab13772).
Werecommend blocking using 2-5% BSA as we have found that use of 5% milk significantly reduces theband intensityfor beta actin. Please refer to the images section for the blocking comparison data.
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.
细胞定位 Cytoplasm cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
ab8226 staining Beta Actin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 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 ab8226 at 5µg/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 red). Nuclear DNA was labelled with DAPI (shown in blue).
All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lane 1 : A431 (Human epidermoid carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human epithelial cell line from embryonic kidney) Whole Cell Lysate
Lane 3 : NIH 3T3 (Mouse embryo fibroblast cell line) Whole Cell Lysate
Lane 4 : PC12 (Rat adrenal gland 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 kDa
This 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 usingGoat 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.
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 5 g/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.
See Abreview
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, 0.1 g/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
Immunofluorescence using ab8226 at 5 g/ml incubated for 1 hour on Rat Colon Cancer cells.
Cells were fixed with ice-cold methanol for5 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.
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, 0.5 g/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 (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : HEK293 (Human epithelial cell line from embryonic kidney) cell lysate
Lane 4 : NIH/3T3 (Mouse embryo fibroblast cell line) 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 cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : HEK293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-mouse IgG at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 10 seconds
Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) with the Prism Ultra Protein Ladder (ab116028) 5 l used. We recommend using our ECL substrate kit (ab65623).
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 cell line from cervix adenocarcinoma) whole cell lysate
Lanes 2 & 5 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lanes 3 & 6 : NIH/3T3 (Mouse embryo 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
发表研究结果有使用 ab8226？请让我们知道，以便我们可以引用本数据表中的参考文章。
ab8226 被引用在 1895 文献中.
Jin P et al. Salidroside inhibits apoptosis and autophagy of cardiomyocyte by regulation of circular RNA hsa_circ_0000064 in cardiac ischemia-reperfusion injury. Gene 767:145075 (2021).PubMed: 32858179 Noback M et al. Post-weaning social isolation increases ?FosB/FosB protein expression in sex-specific patterns in the prelimbic/infralimbic cortex and hippocampus in mice. Neurosci Lett 740:135423 (2021).PubMed: 33069811 Tang M et al. Antileukemic effect of caffeic acid 3,4-dihydroxyphenetyl ester. Evidences for its mechanisms of action. Phytomedicine 80:153383 (2021).PubMed: 33091855 Jiao B et al. Toluene diisocyanate-induced inflammation and airway remodeling involves autophagy in human bronchial epithelial cells. Toxicol In Vitro 70:105040 (2021).PubMed: 33127434 Gao Y et al. LINC00311 promotes cancer stem-like properties by targeting miR-330-5p/TLR4 pathway in human papillary thyroid cancer. Cancer Med 9:1515-1528 (2020).PubMed: 31894666 View all Publications for this productBlocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 5% Temperature: 25 C
Blocking step (agent) for 1 hour(s) and 0 minute(s) Concentration: 5% Temperature: 22 C
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 5% Temperature: 21 C
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 5% Temperature: RT C
Gel Running Conditions Reduced Non-Denaturing (Native) (4–20% Mini-PROTEAN TGX Precast Protein Gel)
Blocking step Odyssey Blocking Buffer (PBS) as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 100% Temperature: 23 C
Blocking step BSA as blocking agent for 45 minute(s) Concentration: 3% Temperature: 25 C
Blocking step Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 100% Temperature: 25 C
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) Concentration: 3% Temperature: 25 C
Blocking step Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 100% Temperature: 25 C
Yes, F-actin is an alternative name for beta-actin, therefore the antibody will cross react. You will need to run the pull down under denaturing conditions in order to avoid pulling down actin with your NMDA receptors. A RIPA buffer would be an ideal buffer to use for this application.
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