Overview

  • Product name
  • Description
    Rabbit polyclonal to SC35
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein (Human)

  • Positive control
    • HeLa nuclear extract

Properties

Applications

Our Abpromise guarantee covers the use of ab28428 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 1/2000 - 1/5000. Detects a band of approximately 30 kDa (predicted molecular weight: 26 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    Necessary for the splicing of pre-mRNA. It is required for formation of the earliest ATP-dependent splicing complex and interacts with spliceosomal components bound to both the 5'- and 3'-splice sites during spliceosome assembly. It also is required for ATP-dependent interactions of both U1 and U2 snRNPs with pre-mRNA. Interacts with other spliceosomal components, via the RS domains, to form a bridge between the 5'- and 3'-splice site binding components, U1 snRNP and U2AF. Binds to purine-rich RNA sequences, either 5'-AGSAGAGTA-3' (S=C or G) or 5'-GTTCGAGTA-3'. Can bind to beta-globin mRNA and commit it to the splicing pathway.
  • Sequence similarities
    Belongs to the splicing factor SR family.
    Contains 1 RRM (RNA recognition motif) domain.
  • Post-translational
    modifications
    Extensively phosphorylated on serine residues in the RS domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 35 kDa antibody
    • arginine/serine-rich 2 antibody
    • PR264 antibody
    • Protein PR264 antibody
    • SC 35 antibody
    • SC-35 antibody
    • SC35 antibody
    • Serine/arginine-rich splicing factor 2 antibody
    • SFRS 2 antibody
    • SFRS2 antibody
    • SFRS2A antibody
    • Splicing component 35 kDa antibody
    • Splicing component antibody
    • Splicing factor antibody
    • Splicing factor arginine/serine rich 2 antibody
    • Splicing factor SC35 antibody
    • Splicing speckle antibody
    • Splicing speckles antibody
    • SR splicing factor 2 antibody
    • SRp30b antibody
    • SRSF2 antibody
    • SRSF2_HUMAN antibody
    see all

Images

  • All lanes : Anti-SC35 antibody (ab28428)

    Lane 1 : recombinant SC35 protein
    Lane 2 : HeLa nuclear lysate

    Predicted band size: 26 kDa

  • IHC image of ab28428 staining in human seminoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab28428, 1µg/ml, 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.

    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.

References

This product has been referenced in:
  • Komeno Y  et al. SRSF2 Is Essential for Hematopoiesis, and Its Myelodysplastic Syndrome-Related Mutations Dysregulate Alternative Pre-mRNA Splicing. Mol Cell Biol 35:3071-82 (2015). Read more (PubMed: 26124281) »
  • Erkelenz S  et al. Balanced splicing at the Tat-specific HIV-1 3'ss A3 is critical for HIV-1 replication. Retrovirology 12:29 (2015). Read more (PubMed: 25889056) »
See all 6 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Bone marrow cell)
Gel Running Conditions
Reduced Denaturing (Any kDMini-PROTEAN TGX Gel)
Loading amount
500000 cells
Specification
Bone marrow cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 10 2015

Question
Answer

Thank you for contacting us.

Your credit note ID is ***


I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone at 888-772-2226. Please refer to the credit note ID in any correspondence with our accounting department.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry these products did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued free of charge replacements with the order number XXXX. These products are expected to arrive to us early next week and we will ship them to you by overnight delivery. These product will the shipped on blue ice as this condition is suitable for the majority of our products.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.

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Answer

Thank you for clarifying this for me.

As you have used these products in applications and species listed on the datasheet and as you have attempted extensive troubleshooting and I can see no further advice which may help in this instance, we believe that the issue may indeed lie in the vials which you have received. It may or may not be shipping or handling related as both products were from the same shipment. Regardless, these products are covered by our Abpromise guarantee. Under this guarantee I may replace or refund these products for you.

While we currently have a new lot of our anti-SC35 antibody (ab28428) available. There is a slight delay with our anti-ZRSR2 antibody as we are awaiting a new lot of this product. The expected arrival of this product is 28 May.

If you would like, I can arrange these replacements to be sent to you free of charge. Please know that these products would also be covered by our Abpromise should you experience any further issues we will replace them or refund the original purchase.

Please let me know how you would like to proceed. Also do not hesitate to contact me with any questions you may have.

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Answer

Thank you for your response.

I do wish to apologize for repeatedly inquiring this further information but I wanted to be clear about the issue you have been having. Based on what you have told me you have previously used the anti-SC35 antibody successfully, in gels loading 25-60ug/well of total protein. During these experiments the product did work. However, you latest experiments using this product in which you have loaded 400ng and 800ng total protein/well and used the product at 1:200 have failed. You have also seen a similar issue when using anti-ZRSR2. Have I got this correct?

Thank you for your help in clarifying this for me. I look forward to your response and helping you to achieve your desired results. Please let me know if you have any questions.

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Answer

Thank you for this further information.

I would like to repeat that I am sorry that these products are not working as expected for you. I see that you have used the anti-SC35 antibody at a much higher concentration than we have had to use in our tests and the anti-ZRSR2 at the recommended starting dilution. My only concern is that you use much less protein than we normally recommend, which is usually ˜25-50ug per well though it may be a low as 10ug. While the levels which you use might work with the purified protein or peptide we are unsure whether there would be enough SC35 or ZRSR2 within your sample for our product to detect. When you were using these products with the higher protein levels were you able to get results?

This product is covered by our Abpromise guarantee. If we cannot remedy this issue together we will replace or refund it under our Abpromise guarantee.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Use our products? Submit an Abreview. Earn rewards!

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Question

Inquiry: We have tried using the Anti-SC35 antibody (a b28428) multiple times in our lab without any success. The antibody was ordered back in April and was delivered the following week (April 25th). It was placed in our -20 freezer when we received it. Increasing amounts of protein have been used in multiple gels, the latest of which was both 400 and 800ng of total protein from 293T and HeLa cells. The proteins were denatured and loaded on to 4-20% Tris-Glycine gels, transferred overnight at 4'C using the Criterion Blotter. The blot has been blocked with 5% non-fat dry milk in TBST at one hour at room temp, as well as overnight. When loading the 800ng, primary was used at 1:200 in TBST 5% milk and incubated overnight at 4'C, washed 3x10 minutes in TBST, incubated with secondary at 25'C for one hour (Donkey Anti-Rabbit IgG HRP from Jackson ImmnuoResearch) at 1:50,000 and in 5% milk TBST, and washed 3x 10minutes in TBST. The blots were then developed with SuperSignal West Femto ECL for five minutes, and then exposed to high-sensitivity x-ray for up to 30 minutes. No bands were seen, however the same samples (and same wash, secondary conditions) had bright, strong bands for two other proteins. Is there any chance this antibody went bad? If so, would it be possible to get a new lot of it? **The same Westerns were also blotted with Anti-ZRSR2 (a b50753), ordered at the same time as the SC35 antibody, also resulting in no bands. The lot number for this antibody was not written down before aliquoting and storage at -20. On the same gel as above, this was used at the suggested 2.5 ug/mL and incubated overnight at 4'C. The same washing, secondary and detection conditions were also used.*** Any help with this would be greatly appreciated.

Read More
Answer

Thank you for contacting us.

I am sorry to hear of the issues which you have been facing when using these products. I would like to thank you for including your protocols details with this inquiry. You state that you have use up to 800ng. Are your samples purified? If not we would recommend attempting whole cell or nuclear lysates with a total protein concentration of 20-50ug/well.

You also mention that other antibodies used on the same blot did work. Could you tell me which antibodies these where and detail the stripping procedure which you used on the blot? Were these other antibodies used before or after the SC35 and ZRSR2 antibodies?



These products are covered by our Abpromise guarantee. If we cannot remedy this issue we will replace or refund the product/s under our Abpromise guarantee, as you are using them according to specifications listed on our datasheet.

I look forward to your response. Please do not hesitate to contact us if you need any advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549 cells (alveolar basal epithelial cells))
Specification
A549 cells (alveolar basal epithelial cells)
Fixative
Formaldehyde
Permeabilization
Yes - 0.5% Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 24 2011

Question
Answer

Thank you for your enquiry. ab28428 is a polyclonal antibody generated from baculovirus-expressed recombinant protein and purified through a protein A column. The size of the protein (SC35) detected from HeLa cell nuclear extract is about 30kDa. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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