Overview

  • Product name

  • Description

    Mouse polyclonal to SRSF5
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Full length protein corresponding to Human SRSF5.

  • Positive control

    • Human liver tissue lysate. Human ovary tissue sections. Hela cells. SRSF5 transfected 293T cell lysate
  • General notes

    Previously labelled as SFRS5.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Constituent: 2.68% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab67175 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 1/500 - 1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 31 kDa).
IHC-P Use a concentration of 1.5 µg/ml.
ICC/IF Use at an assay dependent dilution. NOTE: Product requires purification with Protein A prior to use in ICC/IF and IHC-P.

Target

  • Function

    Plays a role in constitutive splicing and can modulate the selection of alternative splice sites.
  • Sequence similarities

    Belongs to the splicing factor SR family.
    Contains 2 RRM (RNA recognition motif) domains.
  • Post-translational
    modifications

    Extensively phosphorylated on serine residues in the RS domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • arginine/serine-rich 5 antibody
    • Delayed early protein HRS antibody
    • Delayed-early protein HRS antibody
    • HRS antibody
    • Pre mRNA splicing factor SRP40 antibody
    • Pre-mRNA-splicing factor SRP40 antibody
    • Serine and arginine rich splicing factor 5 antibody
    • Serine/arginine-rich splicing factor 5 antibody
    • SFRS 5 antibody
    • SFRS5 antibody
    • Splicing factor antibody
    • Splicing factor arginine/serine rich 5 antibody
    • SRP 40 antibody
    • SRP40 antibody
    • SRSF5 antibody
    • SRSF5_HUMAN antibody
    see all

Images

  • Anti-SRSF5 antibody (ab67175) at 1/500 dilution + human liver tissue lysate at 25 µg/ml

    Secondary
    Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

    Predicted band size: 31 kDa
    Observed band size: 39 kDa
    why is the actual band size different from the predicted?

  • All lanes : Anti-SRSF5 antibody (ab67175) at 1/500 dilution

    Lane 1 : SRSF5 transfected 293T cell lysate
    Lane 2 : Non-transfected 293T cell lysate

    Lysates/proteins at 25 µg/ml per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

    Predicted band size: 31 kDa
    Observed band size: 39 kDa why is the actual band size different from the predicted?
    Additional bands at: 27 kDa. We are unsure as to the identity of these extra bands.

  • ab67175, at 1.5µg/ml, staining SRSF5 in human ovary by immunohistochemistry using formalin-fixed, paraffin-embedded tissue.

  • ICC/IF image of ab67175 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab67175, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:

  • Chen Y  et al. Mutually exclusive acetylation and ubiquitylation of the splicing factor SRSF5 control tumor growth. Nat Commun 9:2464 (2018). Read more (PubMed: 29942010) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (lymphocytes)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
14 µg
Specification
lymphocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 25 2016

Answer

Thank you for your email. The antibody is tested in Hela cells, liver tissue lysates and SFRS5 overexpressing 293T lysates. I haven’t included hela in my last email because we do not have images for comparison. I suggest trying the recommended steps. If in case the results do not improve let me know I will be happy to send a replacement vial from different lot or a different antibody against the same target or I can also provide the ab specific blocking peptide. I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

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Answer

Thank you for your enquiry regarding ab67175 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. The protocol looks fine to me however I would like to make few comments/suggestions that might help to improve your results; - Denature protein for 10 minutes at 100C - Use 5% BSA for blocking - Try loading 10 ug of protein - Use fresh protease inhibitors We use these optimized steps in our own lab with fine results, you may consider trying these. The SRSF5 protein shows 4 different isoforms with molecular weight of 32, 13, and 31 kDa. http://www.uniprot.org/uniprot/Q13243#section_seq. It may well be that the observed molecular weight correspond to isoform that is expressed in cancerous cell line than normal. We have used normal liver lysates as positive control and observed mol wt 39 Kda; I suggest using the same lysates as positive control. We do have these lysates in catalogue the product IDs are ab29889, ab44475, ab44477, ab44479, ab44481 etc. The other option could be trying the blocking I hope these suggestions will help to improve the results. Should you have any other question or need further assistance please do not hesitate to contact us.

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