Rabbit Recombinant Monoclonal alpha skeletal muscle Actin antibody. Carrier free. Suitable for IHC-P, ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
IHC-P | ICC/IF | IP | WB | Flow Cyt (Intra) | |
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Human | Tested | Tested | Expected | Tested | Tested |
Mouse | Tested | Tested | Tested | Tested | Expected |
Rat | Tested | Expected | Expected | Expected | Expected |
Species | Dilution info | Notes |
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Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Mouse, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
ACTA, ACTA1, ACTA, Alpha-actin-1
Rabbit Recombinant Monoclonal alpha skeletal muscle Actin antibody. Carrier free. Suitable for IHC-P, ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
Yes
EPR8484
Affinity purification Protein A
This antibody will detect alpha and gamma specific actin from skeletal, cardiac and smooth muscle.
Blue Ice
+4°C
+4°C
Do Not Freeze
ab224207 is the carrier-free version of Anti-muscle Actin antibody [EPR8484] ab156302.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
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 supplementary information is collated from multiple sources and compiled automatically.
Muscle Actin also known as ACTA1 is a protein that plays a significant role in muscle contraction. It has a molecular weight of approximately 42 kDa. This protein is primarily found in skeletal muscle tissues. Muscle Actin exists in two forms: G-actin a globular monomer and F-actin a filamentous polymer that forms actin filaments. These filaments provide structural support and are involved in muscle contraction through their interaction with myosin.
Muscle Actin interacts with myosin to facilitate muscle contraction. It is an important component of the thin filament within the sarcomere the basic unit of muscle fiber. Muscle Actin is part of a complex and this complex includes other important proteins like tropomyosin and troponin. These proteins regulate the interaction between actin and myosin enabling the muscle contraction process. Muscle Actin's polymerization and depolymerization dynamics are important for the regulation of cellular movement and structural integrity.
Muscle Actin is essential for the actin-myosin contractile machinery in muscle cells which is part of the sarcomere assembly pathway. This process involves other proteins such as myosin heavy chain and tropomyosin which are important for muscle contraction. Another important pathway is the Rho GTPase signaling pathway which controls actin filament assembly and is involved in a variety of cellular processes including muscle contraction and cell motility.
Muscle Actin mutations link to certain muscle-related diseases such as nemaline myopathy and congenital myopathies. Nemaline myopathy involves the presence of rod-like structures in muscle fibers and it results from anomalies in actin filaments. Muscle Actin also associates with cardiac muscle conditions when there are mutations affecting cardiac actin structure or function. This reveals the connection between Muscle Actin tropomyosin and troponin in muscle contraction efficiency and structural integrity contributing to these diseases' pathogenesis.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
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 Alexa Fluor® 488 Anti-muscle Actin antibody [EPR8484] ab190196 for protocol details.
Alexa Fluor® 488 Anti-muscle Actin antibody [EPR8484] 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 Alexa Fluor® 488 Anti-muscle Actin antibody [EPR8484] ab190196 at 1/50 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
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 Alexa Fluor® 647 Anti-muscle Actin antibody [EPR8484] ab190567 for protocol details.
Alexa Fluor® 647 Anti-muscle Actin antibody [EPR8484] 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 Alexa Fluor® 647 Anti-muscle Actin antibody [EPR8484] ab190567 at a 1/50 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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).
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-muscle Actin antibody [EPR8484] (Anti-muscle Actin antibody [EPR8484] ab156302) at 1/10000 dilution
Lane 1: NIH/3T3 whole cell lysate at 10 µg
Lane 2: Rat spleen tissue lysate at 10 µg
Lane 3: L6 whole cell lysate at 10 µg
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 Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-muscle Actin antibody [EPR8484] (Anti-muscle Actin antibody [EPR8484] ab156302) at 1/10000 dilution
All lanes: Human stomach tissue lysate at 10 µg
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 Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.
Anti-muscle Actin antibody [EPR8484] 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 (+): Anti-muscle Actin antibody [EPR8484] ab156302 + NIH/3T3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-muscle Actin antibody [EPR8484] ab156302 in NIH/3T3 whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-muscle Actin antibody [EPR8484] (Anti-muscle Actin antibody [EPR8484] ab156302)
Predicted band size: 42 kDa
Observed band size: 42 kDa
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-muscle Actin antibody [EPR8484] (Anti-muscle Actin antibody [EPR8484] ab156302) at 1/1000 dilution
All lanes: A673 whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa
Blocking and dilution buffer: 5% NFDM /TBST.
This data was developed using Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) 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.
This data was developed using Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) 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.
This data was developed using Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) 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.
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.Immunocytochemistry/Immunofluorescence analysis of A673 cells labelling muscle Actin with purified Anti-muscle Actin antibody [EPR8484] ab156302 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.
Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified Anti-muscle Actin antibody [EPR8484] 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).
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.
All lanes: Western blot - Anti-muscle Actin antibody [EPR8484] (Anti-muscle Actin antibody [EPR8484] ab156302) at 1/1000 dilution
Lane 1: NIH 3T3 lysate at 10 µg
Lane 2: Human fetal artery lysate at 10 µg
Lane 3: Human fetal kidney lysate at 10 µg
Lane 4: Human uterus lysate at 10 µg
Lane 5: Human stomach lysate at 10 µg
Lane 6: Human fetal heart lysate at 10 µg
Lane 7: Human skeletal muscle lysate at 10 µg
Predicted band size: 42 kDa
This data was developed using Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) 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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781).
All lanes: Western blot - Anti-muscle Actin antibody [EPR8484] (Anti-muscle Actin antibody [EPR8484] ab156302) at 1/1000 dilution
Lane 1: A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 2: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg
Lane 3: MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 20 µg
Lane 4: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 5: Skeletal Muscle (Human) Tissue Lysate - adult normal tissue at 20 µg
Lane 6: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (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 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 Anti-muscle Actin antibody [EPR8484] ab156302 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) 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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781).
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.IHC image of unpurified Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] 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.
This data was developed using Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 at a dilution of 1/1000.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
This data was developed using Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 at a dilution of 1/1000.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.Unpurified Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 at 10μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and goat anti-mouse Alexa Fluor® 594 secondary (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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.
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.Unpurified Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302 at 5μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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).
This data was developed using Anti-muscle Actin antibody [EPR8484] ab156302, the same antibody clone in a different buffer formulation.Overlay histogram showing HeLa cells stained with unpurified Anti-muscle Actin antibody [EPR8484] 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 Anti-muscle Actin antibody [EPR8484] ab156302, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor® 488 (IgG H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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.
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
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