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  1. Link

    products/primary-antibodies/smoothelin-antibody-r4a-ab8969.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Intermediate Filaments Class III Smoothelins
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Anti-Smoothelin antibody [R4A] (ab8969)

  • Datasheet
  • SDS
Reviews (5)Q&A (2)References (25)

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Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
  • Western blot - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)
  • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)

Key features and details

  • Mouse monoclonal [R4A] to Smoothelin
  • Suitable for: ICC/IF, IHC-Fr, IHC-P, WB
  • Reacts with: Chicken, Cat, Dog, Human, Pig, Monkey
  • Isotype: IgG1

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Overview

  • Product name

    Anti-Smoothelin antibody [R4A]
    See all Smoothelin primary antibodies
  • Description

    Mouse monoclonal [R4A] to Smoothelin
  • Host species

    Mouse
  • Specificity

    Reacts with the 59 kDa and 100 kDa protein, corresponding to smoothelin A and B, respectively, which are exclusively found in smooth muscle cells.

  • Tested applications

    Suitable for: ICC/IF, IHC-Fr, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Chicken, Cat, Dog, Human, Pig, Monkey
    Does not react with: Mouse, Rat, Hamster
  • Immunogen

    Recombinant full length protein corresponding to Smoothelin. Derived by fusion of SP2/0-Ag14 mouse myeloma cells with spleen cells from a BALB/c mouse immunized with a cytoskeletal extract of chicken gizzard.
    Database link: P53814

  • General notes

    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

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    R4A
  • Myeloma

    Sp2/0-Ag14
  • Isotype

    IgG1
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class III
    • Smoothelins
    • Cardiovascular
    • Vasculature
    • Smooth muscle cell (SMC)
    • Cytoskeleton
    • Developmental Biology
    • Organogenesis
    • Skeletal development
    • Muscle
    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Smooth Muscle Cells

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8969 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
ICC/IF (1)
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
IHC-P (3)
Use at an assay dependent concentration.
WB (1)
1/100 - 1/1000.
Notes
ICC/IF
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
WB
1/100 - 1/1000.

Target

  • Function

    Structural protein of the cytoskeleton.
  • Tissue specificity

    Smooth muscle; contractile or vascular (for the long form).
  • Sequence similarities

    Belongs to the smoothelin family.
    Contains 1 CH (calponin-homology) domain.
  • Cellular localization

    Cytoplasm > cytoskeleton. Exhibits a filamentous organization.
  • Target information above from: UniProt accession P53814 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 373982 Chicken
    • Entrez Gene: 6525 Human
    • Omim: 602127 Human
    • SwissProt: P53814 Human
    • Unigene: 149098 Human
    • Alternative names

      • FLJ35168 antibody
      • FLJ35365 antibody
      • FLJ35620 antibody
      • FLJ38597 antibody
      • Smoothelin antibody
      • SMSMO antibody
      • Smtn antibody
      • SMTN_HUMAN antibody
      see all

    Images

    • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)

      Smooth muscle cells in swine colon as detected by ab8969 (dilution 1:100).

    • Western blot - Anti-Smoothelin antibody [R4A] (ab8969)
      Western blot - Anti-Smoothelin antibody [R4A] (ab8969)
      All lanes : Anti-Smoothelin antibody [R4A] (ab8969)

      Lane 1 : uterine artery
      Lane 2 : uterine corpus.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smoothelin antibody [R4A] (ab8969)

      Immunohistochemical analysis on paraffin section of human smooth muscle labeling Smoothelin with ab8969.

    • Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)

      Indirect immunofluorescence staining of a cultured human fibroblast, showing colocalization of smoothelin (red fluorescence; as detected by ab8969) and actin (green fluorescence).

    • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)

      Smooth muscle cells in swine colon as detected by ab8969 (dilution 1:100).

    • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)

      Smooth muscle cells in swine colon as detected by ab8969 (dilution 1:100).

    • Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)

      Human blood vessel in cardiac muscle stained with the monoclonal smoothelin antibody ab8969.

    • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)

      Immunohistochemical analysis on frozen section of chicken gizzard striated muscle labeling Smoothelin with ab8969.

    • Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunocytochemistry/ Immunofluorescence - Anti-Smoothelin antibody [R4A] (ab8969)This image is courtesy of an anonymous Abreview
      ab8969 staining cells from a human vascular smooth muscle cells by ICC/IF. Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 10% serum for 10 minutes at 21°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 21°C. An Rhodamine conjugated donkey anti-mouse antibody, diluted 1/200, was used as the secondary.

      See Abreview

    • Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)
      Immunohistochemistry (Frozen sections) - Anti-Smoothelin antibody [R4A] (ab8969)

      Immunohistochemistry on frozen section of chicken gizzard striated muscle labeling Smoothelin with ab8969.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (25)

    Publishing research using ab8969? Please let us know so that we can cite the reference in this datasheet.

    ab8969 has been referenced in 25 publications.

    • Xia Y  et al. MIRLET7BHG promotes hepatocellular carcinoma progression by activating hepatic stellate cells through exosomal SMO to trigger Hedgehog pathway. Cell Death Dis 12:326 (2021). PubMed: 33771969
    • Jin YP  et al. Encapsulated three-dimensional bioprinted structure seeded with urothelial cells: a new construction technique for tissue-engineered urinary tract patch. Chin Med J (Engl) 133:424-434 (2020). PubMed: 31977553
    • Ackers I  et al. Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line. Front Cardiovasc Med 7:567837 (2020). PubMed: 33330641
    • Dadhich P  et al. BioSphincters to treat Fecal Incontinence in Nonhuman Primates. Sci Rep 9:18096 (2019). PubMed: 31792260
    • Lau S  et al. Biochemical myogenic differentiation of ASC is donor-dependent and requires sound characterization. Tissue Eng Part A N/A:N/A (2019). PubMed: 30648499
    View all Publications for this product

    Customer reviews and Q&As

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

    1-7 of 7 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smoothelin antibody [R4A]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Sheep Tissue sections (Carotid Artery)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH9.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Carotid Artery
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Le Zhen

    Verified customer

    Submitted Aug 23 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smoothelin antibody [R4A]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rabbit Tissue sections (Carotid Artery)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10 mM Citrate Buffer (pH=6)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    Carotid Artery
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 20 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Smoothelin antibody [R4A]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Vascular smooth muscle cells)
    Specification
    Vascular smooth muscle cells
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% triton X-100
    Blocking step
    Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Mr. G Krenning

    Verified customer

    Submitted Sep 01 2008

    Western blot abreview for Anti-Smoothelin antibody [R4A]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (aortic smooth muscle cells)
    Loading amount
    10 µg
    Specification
    aortic smooth muscle cells
    Gel Running Conditions
    Reduced Denaturing (Tris-acetate 3-8%)
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 01 2008

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smoothelin antibody [R4A]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue lysate - other (canine myocardial infarct)
    Specification
    canine myocardial infarct
    Antigen retrieval step
    None
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Marcin Dobaczewski

    Verified customer

    Submitted Aug 11 2005

    Question

    Hello ThereI have a query regarding an antibody that I received from you. It is Ab8969/ Smoothelin antibody, directed against contractile SMC's.I have been trying to optimize this antibody for sometime now - and the pattern of staining that I am getting is not consistant with smooth muscle cells, BUT rather of Trophoblasts (found in Epithelial tissue). The control tissue that I have been using is placenta & tonsil, both containing several vessels and arteries which should contain smoothelin positive SMC's. However in both controls, only trophoblasts stain up, AND extremely specifically. I can send you an image if that would help you.The pathologists suggest that the antibody in the vial is incorrect, and in fact not anti- Smoothelin at all, since there is absolutely no trace of staining in a single artery or contractile vessel. Is this possible???? This is a really important study we are doing in cardiac research, and we would like to demonstrate smoothelin.I would appreciate your assistance in this rather mystifying situation.Many Thanks

    Read More

    Abcam community

    Verified customer

    Asked on Apr 07 2005

    Answer

    Thank you for your enquiry and your interest in our products. We would like to ask you please to provide some more information about your IHC staining and fill in this specific IHC Questionnaire. Your answer will help us identify the source of the problem and give you sufficient technical support. 1. Order details: • Batch number: • Abcam order or Purchase order number: • Antibody storage conditions (temperature/reconstitution etc) 2. Please describe the problem (high background, no staining etc). 3. On what material are you testing the antibody in IHC • Species: • Cell Iine or tissue: 4. How did you fix the samples? • Ethanol, methanol: (was it ice cold) • Acetone: (was it ice cold) • Paraformaldehyde (percentage, perfusion fixed on not): • Fixation time: • Fixation temperature: 5. For formalin-fixed paraffin embedded tissue: did you apply antigen retrieval step? • Enzymatic method: • Heat mediated technique: • Other : 6. Blocking steps: • For HRP detection method: did you block endogenous peroxidases: • How did you block the unspecific binding sites: serum, BSA, Milk, other • For how long: 7. Primary antibody • Specification (in which species was it raised against): • At what dilution(s) have you tested this antibody: • What dilution buffer was used: • Incubation time: • Incubation temperature: • What washing steps were done: 8. Secondary antibody • Specification (in which species was it raised against): • What dilution buffer was used: • Incubation time: • Incubation temperature: • What washing steps were done: • Do you know whether the problems you are experiencing come from the secondary? 9. What detection method are you using? 10. Background staining • Please provide an image of your staining 11. Which detection system did you use? 12. Did you apply positive and negative controls along with the samples? Please specify. 13. Optimization attempts • How many times have you tried the IHC? • Do you obtain the same results every time? • What steps have you altered? We look forward to hearing from you soon.

    Read More

    Abcam Scientific Support

    Answered on Apr 12 2005

    Question

    Can you tell me the concentration of this ab or reccommendations for dilutions to use for immunohistochemistry?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 28 2003

    Answer

    Ab8969 is in an unpurified form (tissue culture supernatant) and we cannot guarantee the concentrations for unpurified material. When we know what the recommended dilutions are for an antibody we will state them on the datasheet, otherwise the dilutions are assay dependent.

    Read More

    Abcam Scientific Support

    Answered on Oct 28 2003

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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