muscle Actin重组抗体[EPR8484]_muscle Actin抗体(ab224207)

产品名称

Anti-muscle Actin抗体[EPR8484] - BSA and Azide free
参阅全部 muscle Actin 一抗
描述

兔单克隆抗体[EPR8484] to muscle Actin - BSA and Azide free
宿主

Rabbit
特异性

This antibody will detect alpha and gamma specific actin from skeletal, cardiac and smooth muscle.
经测试应用

适用于: ICC/IF, IHC-P, Flow Cyt (Intra), IP, WBmore details
种属反应性

与反应: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
常规说明
ab224207 is the carrier-free version of ab156302.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C. Do Not Freeze.
存储溶液

pH: 7.2
Constituent: PBS
无载体

是
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR8484
同种型

IgG
研究领域

The Abpromise guarantee

Abpromise™承诺保证使用ab224207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。

应用	Ab评论	说明
ICC/IF		Use at an assay dependent concentration.
IHC-P		Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
Flow Cyt (Intra)		Use at an assay dependent concentration.
IP		Use at an assay dependent concentration.
WB		Use at an assay dependent concentration. Predicted molecular weight: 42 kDa.

说明
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
Flow Cyt (Intra) Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 42 kDa.

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 ACTA1 are the cause of nemaline myopathy type 3 (NEM3) [MIM:161800]. A form of nemaline myopathy. Nemaline myopathies are muscular disorders characterized by muscle weakness of varying severity and onset, and abnormal thread-or rod-like structures in muscle fibers on histologic examination. The phenotype at histological level is variable. Some patients present areas devoid of oxidative activity containg (cores) within myofibers. Core lesions are unstructured and poorly circumscribed.
Defects in ACTA1 are a cause of myopathy, actin, congenital, with excess of thin myofilaments (MPCETM) [MIM:161800]. A congenital muscular disorder characterized at histological level by areas of sarcoplasm devoid of normal myofibrils and mitochondria, and replaced with dense masses of thin filaments. Central cores, rods, ragged red fibers, and necrosis are absent.
Defects in ACTA1 are a cause of congenital myopathy with fiber-type disproportion (CFTD) [MIM:255310]; also known as congenital fiber-type disproportion myopathy (CFTDM). CFTD is a genetically heterogeneous disorder in which there is relative hypotrophy of type 1 muscle fibers compared to type 2 fibers on skeletal muscle biopsy. However, these findings are not specific and can be found in many different myopathic and neuropathic conditions.
序列相似性

Belongs to the actin family.
翻译后修饰

Oxidation of Met-46 by MICALs (MICAL1, MICAL2 or MICAL3) to form methionine sulfoxide promotes actin filament depolymerization. Methionine sulfoxide is produced stereospecifically, but it is not known whether the (S)-S-oxide or the (R)-S-oxide is produced.
细胞定位

Cytoplasm > cytoskeleton.
Target information above from: UniProt accession P68133 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 58 Human
- Entrez Gene: 11459 Mouse
- Entrez Gene: 29437 Rat
- Omim: 102610 Human
- SwissProt: P68133 Human
- SwissProt: P68033 Mouse
- SwissProt: P68134 Mouse
- SwissProt: P68136 Rat
- Unigene: 1288 Human
- Unigene: 214950 Mouse
- Unigene: 82732 Rat
see all
别名
- a actin antibody
- ACTA antibody
- ACTA1 antibody
- ACTA2 antibody
- ACTC antibody
- ACTC1 antibody
- Actin antibody
- ACTS_HUMAN antibody
- ACTSA antibody
- Alpha 2 actin antibody
- alpha skeletal muscle antibody
- Alpha-actin-1 antibody
- Cardiac muscle alpha actin 1 antibody
- Skeletal muscle alpha actin 1 antibody
see all

Western blot - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

All lanes : Anti-muscle Actin antibody [EPR8484] (ab156302) at 1/10000 dilution (purified)

Lane 1 : NIH/3T3 whole cell lysate
Lane 2 : Rat spleen tissue lysate
Lane 3 : L6 whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG (specific to the non-reduced form of IgG) at 1/10000 dilution

Predicted band size: 42 kDa
Observed band size: 42 kDa

This data was developed using ab156302, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Flow Cytometry (Intracellular) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.

Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab156302 at a dilution of 1 in 50 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
Western blot - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

Anti-muscle Actin antibody [EPR8484] (ab156302) at 1/10000 dilution (purified) + Human stomach tissue lysate at 10 µg

Secondary
HRP-conjugated goat anti-rabbit IgG (specific to the non-reduced form of IgG) at 1/10000 dilution

Predicted band size: 42 kDa
Observed band size: 42 kDa

This data was developed using ab156302, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

Anti-muscle Actin antibody [EPR8484] (ab156302) at 1/1000 dilution (purified) + A673 whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 42 kDa
Observed band size: 42 kDa

This data was developed using ab156302, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat colon tissue labelling muscle Actin with purified ab156302 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue labelling muscle Actin with purified ab156302 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling muscle Actin with purified ab156302 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Immunocytochemistry/Immunofluorescence analysis of A673 cells labelling muscle Actin with purified ab156302 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Flow Cytometry (Intracellular) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.

Intracellular Flow Cytometry analysis of HeLa cells labelling muscle Actin with purified ab156302 at 1/50 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunoprecipitation - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.

ab156302 (purified) at 1/20 immunoprecipitating muscle Actin in NIH/3T3 whole cell lysate.

Lane 1 (input): NIH/3T3 whole cell lysate (10µg)

Lane 2 (+): ab156302 + NIH/3T3 whole cell lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab156302 in NIH/3T3 whole cell lysate.

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

All lanes : Anti-muscle Actin antibody [EPR8484] (ab156302) at 1/1000 dilution (unpurified)

Lane 1 : NIH 3T3 lysate
Lane 2 : Human fetal artery lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human uterus lysate
Lane 5 : Human stomach lysate
Lane 6 : Human fetal heart lysate
Lane 7 : Human skeletal muscle lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 42 kDa

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Western blot - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

All lanes : Anti-muscle Actin antibody [EPR8484] (ab156302) at 1/1000 dilution (unpurified)

Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue
Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

Predicted band size: 42 kDa
Observed band size: 42 kDa

This data was developed using ab156302, the same antibody clone in a different buffer formulation.

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 Licor blocking buffer before being incubated with ab156302 overnight at 4°C. Antibody binding was detected using ab175781 at a 1/10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

Secondary antibody - goat anti-rabbit Alexa Fluor^® 790 (ab175781).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
IHC image of unpurified ab156302 staining muscle 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 unpurified ab156302, 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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue labelling muscle Actin with unpurified ab156302 at a dilution of 1/1000. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human heart muscle tissue labelling muscle Actin with unpurified ab156302 at a dilution of 1/1000. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.
Unpurified ab156302 staining Actin in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with unpurified ab156302 at 10μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an goat anti-rabbit Alexa Fluor^® 488 secondary (ab150081) at 2 μg/ml (shown in green) and goat anti-mouse Alexa Fluor^® 594 secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.Negative controls: 1 – Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
Immunocytochemistry/ Immunofluorescence - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.Unpurified ab156302 staining Actin in NIH3T3 cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with unpurified ab156302 at 5μg/ml and ab195889 at 1/250 dilution (shown in pseudo color red) overnight at +4°C, followed by a further incubation at room temperature for 1h with an goat anti-rabbit Alexa Fluor^® 488 secondary (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

Clone EPR8484 (ab224207) has been successfully conjugated by Abcam. This image was generated using Anti-muscle Actin antibody [EPR8484] (Alexa Fluor® 488). Please refer to ab190196 for protocol details.
ab190196 staining Actin in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab190196 at 1/50 dilution and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat anti-mouse AlexaFluor^®594 (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Flow Cytometry (Intracellular) - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.Overlay histogram showing HeLa cells stained with unpurified ab156302 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab156302, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor^® 488 (IgG H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Immunocytochemistry/ Immunofluorescence - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

Clone EPR8484 (ab224207) has been successfully conjugated by Abcam. This image was generated using Anti-muscle Actin antibody [EPR8484] (Alexa Fluor® 647). Please refer to ab190567 for protocol details.
ab190567 staining Actin in NIH3T3 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% 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 ab190567 at a 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in NIH3T3 cells fixed with 4% formaldehyde (10 min).
OI-RD Scanning - Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

This data was developed using ab156302, the same antibody clone in a different buffer formulation.Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
Anti-muscle Actin antibody [EPR8484] - BSA and Azide free (ab224207)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheet download

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Key features and details

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概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

常规说明

性能

形式

存放说明

存储溶液

无载体

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

Related Products

应用

The Abpromise guarantee

靶标

功能

疾病相关

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

Datasheet download

Certificate of Compliance

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