Overview

  • Product name
  • Description
    Rabbit polyclonal to Sumo 1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Arabidopsis thaliana
  • Immunogen

    Recombinant full length protein (Arabidopsis thaliana).

  • Positive control
    • Arabidopsis extracts that have been heat treated for for 30-120 mins by moving them from 24 to 37 degrees C. Recombinant SUMO1 is detectable at concentrations as low as 5ng/ml.

Properties

Applications

Our Abpromise guarantee covers the use of ab5316 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. Predicted molecular weight: 12 kDa.

Target

  • Function
    Ubiquitin-like protein that can be covalently attached to proteins as a monomer or a lysine-linked polymer. Covalent attachment via an isopeptide bond to its substrates requires prior activation by the E1 complex SAE1-SAE2 and linkage to the E2 enzyme UBE2I, and can be promoted by E3 ligases such as PIAS1-4, RANBP2 or CBX4. This post-translational modification on lysine residues of proteins plays a crucial role in a number of cellular processes such as nuclear transport, DNA replication and repair, mitosis and signal transduction. Involved for instance in targeting RANGAP1 to the nuclear pore complex protein RANBP2. Polymeric SUMO1 chains are also susceptible to polyubiquitination which functions as a signal for proteasomal degradation of modified proteins. May also regulate a network of genes involved in palate development.
  • Involvement in disease
    Defects in SUMO1 are the cause of non-syndromic orofacial cleft type 10 (OFC10) [MIM:613705]; also called non-syndromic cleft lip with or without cleft palate 10. OFC10 is a birth defect consisting of cleft lips with or without cleft palate. Cleft lips are associated with cleft palate in two-third of cases. A cleft lip can occur on one or both sides and range in severity from a simple notch in the upper lip to a complete opening in the lip extending into the floor of the nostril and involving the upper gum. Note=A chromosomal aberation involving SUMO1 is the cause of OFC10. Translocation t(2;8)(q33.1;q24.3). The breakpoint occurred in the SUMO1 gene and resulted in haploinsufficiency confirmed by protein assays.
  • Sequence similarities
    Belongs to the ubiquitin family. SUMO subfamily.
    Contains 1 ubiquitin-like domain.
  • Post-translational
    modifications
    Cleavage of precursor form by SENP1 or SENP2 is necessary for function.
    Polymeric SUMO1 chains undergo polyubiquitination by RNF4.
  • Cellular localization
    Nucleus membrane. Nucleus speckle. Cytoplasm. Recruited by BCL11A into the nuclear body.
  • Information by UniProt
  • Alternative names
    • DAP1 antibody
    • GAP modifying protein 1 antibody
    • GAP-modifying protein 1 antibody
    • GMP 1 antibody
    • GMP1 antibody
    • OFC10 antibody
    • PIC 1 antibody
    • PIC1 antibody
    • SENP2 antibody
    • Sentrin 1 antibody
    • Sentrin antibody
    • Small ubiquitin related modifier 1 antibody
    • Small ubiquitin-like modifier 1 antibody
    • Small ubiquitin-related modifier 1 antibody
    • SMT3 antibody
    • SMT3 homolog 3 antibody
    • SMT3 suppressor of mif two 3 homolog 1 antibody
    • SMT3, yeast, homolog 3 antibody
    • Smt3C antibody
    • SMT3H3 antibody
    • SUMO-1 antibody
    • SUMO1 antibody
    • SUMO1_HUMAN antibody
    • Ubiquitin homology domain protein PIC1 antibody
    • Ubiquitin Like 1 antibody
    • Ubiquitin like protein SMT3C antibody
    • Ubiquitin like protein UBL1 antibody
    • Ubiquitin-homology domain protein PIC1 antibody
    • Ubiquitin-like protein SMT3C antibody
    • Ubiquitin-like protein UBL1 antibody
    • UBL 1 antibody
    • UBL1 antibody
    see all

References

This product has been referenced in:
See all 18 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Plants Tissue lysate - whole (anther)
Loading amount
30 µg
Specification
anther
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Dec 05 2012

Answer

Thank you for your phone call and I'm sorry to hear that you are experiencing difficulty with ab5316. The replacement vials are being sent to your attention free of charge on order reference # 109485. The item is currently out of stock and so you should receive it early next week. You will receive emails regarding shipping to keep you updated. Please let me know how the replacement works out for you and if you have any additional questions.

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Answer

Thank you for getting back to me and for providing further information about your experiments. As we understand from your recent e-mail, after blocking the membrane (without running the samples) the primary antibody provided a very high background signal. Have you tested the secondary antibody alone (run a secondary control only) to see if this problem really comes from the primary or the secondary antibody? I have checked your order record and the delivery details in our system. Looking at FedEx website I can see that the package was held up at the customs for several days. It may well be that the antibody has just gone off during long delivery. We have the same batch in stock at the moment but I can send you a new vial from the same Lot (free of charge) if you wish to test it. Please get back to me and do let me know how to process. I am looking forward to hearing from you soon.

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Question

1. Order details: •Batch number:lot 25213 •Abcam order or Purchase order number: order ref no. 81493 •Antibody storage conditions (temperature/reconstitution etc): 2. Please describe the problem (high background, wrong band size, more bands, no band etc). very high background. I have troubleshoot by using empty membranes (PVDF and nitrocelullose) without any protein samples, the signal is so high that the membrane glow in the dark. 3. On what material are you testing the antibody in WB? •Species: Aspergillus nidulans •Cell extract or Nuclear extract: Crude extract •Purified protein or Recombinant protein: 3. The lysate •How much protein was loaded: 50ug •What lysis buffer was used: Tris, SDS •What protease inhibitors were used: none •What loading buffer was used: Lammeli •Did you heat the samples: temperature and time: yes, boil for 3mins 4. Electrophoresis/Gel conditions/ Transfer conditions •Reducing or non reducing gel: reducing •Gel percentage : 8% •Transfer conditions: semi-dry 5. Blocking conditions •Buffer:TBS-T •Blocking agent: milk, BSA, serum, what percentage: Tried 5%, 10% milk, 5% BSA •Incubation time:for a few hours to overnight •Incubation temperature:25oC 6. Primary Antibody •Specification (in which species was it raised against): Rabbit •At what dilution(s) have you tested this antibody: 1:1000 as recommended •What dilution buffer was used:5%milk in TBS-T •Incubation time:1hr at 25oC or O/N at 4oC •Incubation temperature:4 and 25 oC tested •What washing steps were done:3x 10 mins with TBS-T 7. Secondary Antibody •Specification (in which species was it raised against)? Goat •At what dilution(s) have you tested this antibody:1/5000 •Incubation time 1hr at 4oC •Wash steps:3x10mins with TBS-T •Do you know whether the problems you are experiencing come from the secondary? It is working fine with my other westerns 8. Detection method ECl, ECl+, other detection method: ECL+ 9. Background bands •Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): other westerns work fine •Is the blocking step sufficient? yes (10% milk overnight) •Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) I have tried that too... •At what size are the bands migrating? Could they be degradation products of your target? empty membrane has high background too •Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 10. Did you apply positive and negative controls along with the samples? Please specify. 11. Optimization attempts •How many times have you tried the Western? at least 5 times •Do you obtain the same results every time e.g. are background bands always in the same place? background everywhere even with empty membrane •What steps have you altered? empty (no protein) nitrocelluse and PVDF, respectively

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Answer

Thank you for contacting us and for taking your time to fill in the Questionnaire and for providing more information about your Western blot assay. This antibody was raised against a recombinant full length protein from Arabidopsis thaliana and it has been characterized using samples from this species. Unfortunately, other species have not been tested yet and we do not know if ab5316 recognizes Sumo 1 in Aspergillus nidulans or not. Here we would like to make some suggestions which could improve the detection: As we understand from your e-mail, you do not use any proteinase inhibitors in the lysis buffer when preparing the samples. It is very important to apply a cocktail of different proteinases in order to prevent the degradation of the target. It is difficult to tell what may have happened in your samples without seeing the Wb image but it may well be that the high background due to protein degradation. You loaded 50 ug of protein onto the gel which is a bit too much. We usually advise 20-30 ug per lane, otherwise the gel is simply overloaded. The excepted band size is 12 kDa. In order to be able to detect a band at such a low MW level, you need to apply high percentage of gel i.e. 15%. If you use 8%, there is a very high chance that the separated target protein has run out of the gel. We hope that this information will be useful for you.

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