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Signal Transduction Cytoskeleton / ECM Cytoskeleton Motor Proteins Myosin
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RecombinantRabMAb

Recombinant Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

  • Datasheet
  • SDS
Reviews (5)Q&A (7)References (25)

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Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • OI-RD Scanning - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
  • Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3741] to Myosin Light Chain 2
  • Suitable for: WB, IHC-P
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-Myosin Light Chain 2 antibody [EPR3741]
    See all Myosin Light Chain 2 primary antibodies
  • Description

    Rabbit monoclonal [EPR3741] to Myosin Light Chain 2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Myosin Light Chain 2 aa 150-250. The exact sequence is proprietary.

  • Positive control

    • WB: Recombinant Human Myosin Light Chain 2 protein (ab117178), Fetal heart, fetal skeletal muscle, Human heart, mouse heart and rat heart lysates IHC-P: Human heart muscle
  • General notes

     

     

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 2.78 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR3741
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Motor Proteins
    • Myosin
    • Cardiovascular
    • Heart
    • Contractility
    • Contractile Proteins
    • Myosins

Associated products

  • Alternative Versions

    • Anti-Myosin Light Chain 2 antibody [EPR3741] - BSA and Azide free (ab183490)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Myosin Light Chain 2 protein (ab79185)

Applications

Our Abpromise guarantee covers the use of ab92721 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/5000. Predicted molecular weight: 19 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The use of an HRP/AP polymerized secondary antibody is recommended.

  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Involvement in disease

      Defects in MYL2 are the cause of cardiomyopathy familial hypertrophic type 10 (CMH10) [MIM:608758]. Familial hypertrophic cardiomyopathy is a hereditary heart disorder characterized by ventricular hypertrophy, which is usually asymmetric and often involves the interventricular septum. The symptoms include dyspnea, syncope, collapse, palpitations, and chest pain. They can be readily provoked by exercise. The disorder has inter- and intrafamilial variability ranging from benign to malignant forms with high risk of cardiac failure and sudden cardiac death.
      Defects in MYL2 are the cause of cardiomyopathy familial hypertrophic with mid-left ventricular chamber type 2 (MVC2) [MIM:608758]. MVC2 is a very rare variant of familial hypertrophic cardiomyopathy, characterized by mid-left ventricular chamber thickening.
    • Sequence similarities

      Contains 3 EF-hand domains.
    • Post-translational
      modifications

      N-terminus is methylated by METTL11A/NTM1.
    • Target information above from: UniProt accession P10916 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 4633 Human
      • Entrez Gene: 17906 Mouse
      • Entrez Gene: 363925 Rat
      • Omim: 160781 Human
      • SwissProt: P10916 Human
      • SwissProt: P51667 Mouse
      • SwissProt: P08733 Rat
      • Unigene: 75535 Human
      • Unigene: 1529 Mouse
      • Unigene: 37176 Rat
      • Unigene: 6534 Rat
      see all
    • Alternative names

      • Cardiac myosin light chain-2 antibody
      • Cardiac ventricular myosin light chain 2 antibody
      • CMH10 antibody
      • MLC 2v antibody
      • MLC-2 antibody
      • MLC-2v antibody
      • MLC2 antibody
      • MLRV_HUMAN antibody
      • MYL 2 antibody
      • MYL2 antibody
      • Myosin light chain 2 regulatory cardiac slow antibody
      • Myosin light polypeptide 2 regulatory cardiac slow antibody
      • Myosin regulatory light chain 2 antibody
      • Myosin regulatory light chain 2 ventricular/cardiac muscle isoform antibody
      • Regulatory light chain of myosin antibody
      • RLC of myosin antibody
      • Slow cardiac myosin regulatory light chain 2 antibody
      • ventricular/cardiac muscle isoform antibody
      see all

    Images

    • Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      All lanes : Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721) at 1/1000 dilution

      Lane 1 : Human fetal heart tissue lysate
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 3 : 293T (Human embryonic kidney epithelial cell), whole cell lysate
      Lane 4 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 19 kDa
      Observed band size: 19 kDa


      Exposure time: 10 seconds


      Blocking/Diluting buffer and concentration 5% NFDM/TBST

      Myosin light chain-2 (MYL2, also called MLC-2) is an ~19kDa sarcomeric protein that belongs to the EF-hand calcium binding protein superfamily and exists as three major isoforms encoded by three distinct genes in mammalian striated muscle (PMID: 26074085). Although some cell lines are reported to be MYL2 positive, such as HeLa, 293T and MCF7 (PMID: 21487036, 21487036, 23093227), ab92721 only works in muscle and heart tissues according to our internal testing.

    • Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      All lanes : Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721) at 1/10000 dilution

      Lane 1 : Fetal heart lysate
      Lane 2 : Fetal skeletal muscle lysate
      Lane 3 : Human heart lysate
      Lane 4 : Mouse heart lysate
      Lane 5 : Rat heart lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size: 19 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721, at a 1/100 dilution, staining Myosin Light Chain 2 in paraffin embedded Human heart muscle by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Western blot - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      All lanes : Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721) at 1/10000 dilution

      Lane 1 : Recombinant Human Myosin Light Chain 2 protein (ab79185) at 0.1 µg
      Lane 2 : Recombinant Human Myosin Light Chain 2 protein (ab79185) at 0.01 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 19 kDa


      Exposure time: 4 minutes
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721 showing positive staining in human Heart muscle tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721 showing positive staining in human Skeletal muscle tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721 showing negative staining in Normal human tonsil tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721 showing negative staining in Normal human brain tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721 showing negative staining in Normal human liver tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721 showing negative staining in Normal human kidney tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

      ab92721 showing negative staining in Normal human breast tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • OI-RD Scanning - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      OI-RD Scanning - Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)
      Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

    Protocols

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (25)

    Publishing research using ab92721? Please let us know so that we can cite the reference in this datasheet.

    ab92721 has been referenced in 25 publications.

    • Guo C  et al. Bilobalide reversibly modulates blood-brain barrier permeability through promoting adenosine A1 receptor-mediated phosphorylation of actin-binding proteins. Biochem Biophys Res Commun 526:1077-1084 (2020). PubMed: 32312522
    • Teigen LE  et al. Ca2+ dependency of limb muscle fiber contractile mechanics in young and older adults. Am J Physiol Cell Physiol 318:C1238-C1251 (2020). PubMed: 32348175
    • Liu Z  et al. Novel circRNA_0071196/miRNA-19b-3p/CIT axis is associated with proliferation and migration of bladder cancer. Int J Oncol 57:767-779 (2020). PubMed: 32705161
    • Hodatsu A  et al. Impact of cardiac myosin light chain kinase gene mutation on development of dilated cardiomyopathy. ESC Heart Fail 6:406-415 (2019). PubMed: 30690923
    • Søgaard PP  et al. Epithelial polarization in 3D matrix requires DDR1 signaling to regulate actomyosin contractility. Life Sci Alliance 2:N/A (2019). PubMed: 30760555
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 12 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Myosin Light Chain 2 antibody [EPR3741]

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% TritonX100 in PBS
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Oct 27 2015

    Western blot abreview for Anti-Myosin Light Chain 2 antibody [EPR3741]

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Ileon)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    15 µg
    Specification
    Ileon
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 30 2015

    Western blot

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15)
    Sample
    Rat Tissue lysate - whole (Skeletal muscle)
    Specification
    Skeletal muscle
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 22 2013

    Western blot

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15)
    Sample
    Mouse Tissue lysate - whole (Heart)
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 22 2013

    Western blot

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15)
    Sample
    Human Tissue lysate - whole (Heart)
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 22 2013

    Question

    I need to order antibodies for MYL7 ((ab92721) and MYL2 (ab137063). While going through the datasheets, I've seen IHC images of these antibodies but no reference is given for them. Can you provide us with the references for those images. Also, as we are regular customers of your products, can you provide us a small gift sample of these antibodies as our project is in infancy at present.

    I look forward for your early response.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 04 2012

    Answer

    Thank you for contacting us with your questions.

    The IHC images on the datasheets for ab92721 and ab137063 were produced in the Epitomics lab, and they weren't taken from publications so there are no references. Ab92721 was used in one publication-

    http://www.ncbi.nlm.nih.gov/pubmed/22037191?dopt=Abstract

    We only have full size vials of our antibodies, so we don't have samples to send out. However, we do have a promotion on rabbit monoclonal antibodies that is valid until December 16. For this promotion, if you purchase any primary antibody you can receive a free rabbit monoclonal. Since both of these antibodies are rabbit monoclonals (also called RabMabs), you could purchase one and then receive the other for free. If you'd like to take advantage of this offer, please include the code RABMAB-XBSMG with your order.

    I hope that this information will be useful, but please let me know if you have further questions or need anything else and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Dec 04 2012

    Question

    Thank you very much for helping me solving this problem.
    The antibody for troponin T that I worked before was ab8295, probably works better with my type of samples.
    I need antibodies that prove cardiac differentiation, that is why I bought cardiac troponin T and MLC2v. If you have suggestions please let me know, so we can make an agreement concerning the replacement of antibodies.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 02 2012

    Answer

    Are you interested in an antibody against a different cardiac marker such as GATA4? The antibody ab86371 looks good.

    Click here (or use the following: https://www.abcam.com/GATA4-antibody-6H10-ab86371.html).

    The antibody ab8295, which you used before successfully, is a cardiac troponin T antibody. Do you want another vial of that, to replace the other troponin T antibody, ab33589? If not, then I suggest ab89594.

    Click here (or use the following: https://www.abcam.com/Myosin-Light-Chain-2-antibody-AT3B2-ab89594.html).

    I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Apr 02 2012

    Question

    I bought 2 antobodies from your company and I am not able to get signal when I perform western-blotting under the conditions advised by you. I tried to increase also the concentration of antibody and I was unable to have a result. I am sure that is not a problem with the secondary antibody because the secondary antibodies worked with other primary antibodies. Both antibodies that I bought from you were shiped together, I don't know if there was some problem with it.
    The antibodies are ab33589 and ab92721. Is is possible to send me other antibodies to replace this ones?
    Thank you very much.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 29 2012

    Answer

    Thank you for bringing this to our attention. If I cannot offer a useful suggestion to improve your results, and if the antibodies are covered by our guarantee, I will send replacements.

    I will first need a few details of your experiment and your order.

    1. Can you please tell me when you ordered the antibodies? If you have a purchase order number or the name of the person who placed the order, that will help me find it in our records.

    2. Please describe your samples: what is the species and cell type or tissue, and how were the lysates prepared? Did you use protease inhibitors?

    3. How much protein did you load per lane of the gel?

    4. Are you confident that the transfer from gel to blot was successful?

    5. What is the blocking solution? (For example, 5% milk in TBS or 1% BSA, etc.).

    6. You mentioned using the antibodies as advised on the datasheets. What dilutions did you try for each antibody?

    7. Are confident the secondary antibody is effective?

    I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Mar 29 2012

    Question

    I have used ab48003 to stain primary mouse cell cultures of cell types I expect to be positive and cell types I expect to be negative for for myosin light chain 2. However all samples stain equally well with this antibody, leading me to think it is reacting non specifically.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 15 2011

    Answer

    I have issued a free of charge replacement for one vial of ab92721. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

    Read More

    Abcam Scientific Support

    Answered on Dec 15 2011

    Question

    What is the WB protocol used for this antibody?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 14 2011

    Answer

    Thank you for contacting Abcam regarding ab92721 and ab89594. I have received the following protocol information from the laboratory: Western Blotting Protocol Solutions and Reagents 2X Laemmli Sample Buffer: 62.5 mM Tris-HCl (pH 6.8), 25% glycerol, 2% SDS, 0.01% Bromophenol Blue, 710 mM beta-mercaptoethanol TBST (50 mM Tris, pH 7.6, 150 mM NaCl, 0.1% Tween-20) TBST is Tris Base Saline buffer with 0.1% Tween-20 5% bovine serum albumin (BSA): Use 25 g in 500 ml TBST Goat anti-rabbit HRP antibody - suggest ab6721 Goat anti-mouse-HRP - suggest ab6789 Chemiluminesence reagents; such as ECL materials and film or imaging system for detection Protocol Run SDS-PAGE using 10-60ug heart lysate and transfer to nitrocellulose membrane. Block nitrocellulose for 1 hour at room temperature or overnight at 4°C using 5% BSA Incubate nitrocellulose with appropriate dilutions of primary antibody in 5% BSA overnight at 4°C or for 2 hours at room temperature Wash nitrocellulose with three 5-min washes using TBST. Incubate nitrocellulose with goat anti-rabbit (or goat anti-mouse) HRP antibody, diluted to 1:500 to 100,000 in 5% BSA for 1-2 hours at room temperature. Wash nitrocellulose in 3 washes of TBST, then rinse in TBS prior to addition of chemiluminesence reagents. Remove excess chemiluminescence reagent and sandwich nitrocellulose blot in any type of transparent plastic wrap (plastic cling wrap, transparent sheet protector, etc.). Acquire image using darkroom development techniques. I hope this information is helpful.  Please to not hesitate to contact us if you have any additional questions.

    Read More

    Abcam Scientific Support

    Answered on Nov 14 2011

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