Overview

  • Product name

  • Description

    Rabbit polyclonal to PURA
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Human
    Predicted to work with: Sheep
  • Immunogen

    Synthetic peptide corresponding to Human PURA aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab88160)

  • Positive control

    • This antibody gave a positive signal in Human Brain Tissue Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab79936 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 Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 34 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function

    This is a probable transcription activator that specifically binds the purine-rich single strand of the PUR element located upstream of the MYC gene. May play a role in the initiation of DNA replication and in recombination.
  • Sequence similarities

    Belongs to the PUR DNA-binding protein family.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • PUR ALPHA antibody
    • PUR1 antibody
    • purA antibody
    • PURA_HUMAN antibody
    • PURALPHA antibody
    • Purine rich element binding protein A antibody
    • Purine rich single stranded DNA binding protein alpha antibody
    • Purine-rich single-stranded DNA-binding protein alpha antibody
    • Transcriptional activator protein PUR alpha antibody
    • Transcriptional activator protein Pur-alpha antibody
    see all

Images

  • Anti-PURA antibody (ab79936) at 1 µg/ml + Human brain tissue lysate - total protein (ab29466) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 34 kDa
    Observed band size: 42 kDa
    why is the actual band size different from the predicted?



    PURA contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. Furthermore, other commercially available antibodies react with a target at approximately 42 kDa.

  • ICC/IF image of ab79936 stained HeLa cells. The cells were 100% Methanol fixed (5 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 (ab79936, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit 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. This antibody also gave a positive result in 100% Methanol fixed (5 min) Hek293. HepG2, MCF-7 cells at 5µg/ml.

References

This product has been referenced in:

  • Swinnen B  et al. A zebrafish model for C9orf72 ALS reveals RNA toxicity as a pathogenic mechanism. Acta Neuropathol 135:427-443 (2018). Read more (PubMed: 29302778) »
  • Moldovan JB & Moran JV The Zinc-Finger Antiviral Protein ZAP Inhibits LINE and Alu Retrotransposition. PLoS Genet 11:e1005121 (2015). Read more (PubMed: 25951186) »
See all 7 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Sample
Mouse Cell lysate - whole cell (whole brain)
Specification
whole brain
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jul 10 2013

Question
Answer

Thank you for contacting us. I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways: (1) Redeemed against the original invoice if this hasn't already been paid. (2) Held on the account for use against a future order. (3) A full refund can be offered where no other invoices are outstanding. Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge. To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website. The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB. 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  

Read More

Question

I recently purchased two antibodies from you and was very disappointed with how they performed. It was mouse anti CoxIV (ab14744) and mouse anti PurA (ab77734) antibodies. They were intended to be used for ICC/IF, but both gave very poor staining.   I have tried the mouse anti CoxIV antibody (ab14744) in both mouse and in human cells. The cells were fixed in 4% formaldehyde and I titrated the antibody dilution between 1:100 and 1:5000, and compared with your rabbit polyclonal to CoxIV antibody (ab16056). ab14744 gave virtually no staining while ab16056 gave very nice mitochondrial staining at 1:1000 dilution. It clearly states in the datasheet that ab14744 should work in ICC/IF and be reactive in both human and mouse. This antibody was mainly intended to be used as a control when running in situ proximity ligation assay (the Duolink kit from Olink), which I mainly use on mES cells. Do you have any other mouse antibodies against any mitochondrial protein that is guaranteed to work in mouse cells for ICC/IF following formaldehyde fixation? Due to the poor performance of the above mentioned antibody I expect a steep discount on any such antibody.   My experience with the mouse anti PurA (ab77734) antibody is similar, but I only bought this antibody because I received a “Telomere antibodies” newsletter from you saying that it had been tested successfully in mouse on ICC/IF. In my hands however it gives no staining at all above background in mES cells, where on the contrary your rabbit anti PurA antibody (ab79936) gives a very strong nuclear staining mainly near the nuclear envelope.   I have been very pleased with your products and service in the past, but was very disappointed with the above mentioned products and thus hope to be fairly compensated.    

Read More
Answer

Thank you for contacting Abcam. I am sorry that you have been experiencing difficulties with our two antibodies, ab14744 and ab77734.  We have received positive feedback for both of these antibodies in ICC and so I am not sure why they have not worked well in your experiments.  I have checked our catalog, and unfortunately these are the only two mouse monoclonal antibodies against these targets.  I am happy to offer replacements, free of charge with a new vial of these antibodies or provide a credit for the products.  Please let me know which you would prefer. I hope this information is helpful and I do apologize for the inconvenience this issue has caused.  Please do not hesitate to contact us if you have any additional questions.  I look forward to your reply so that I may assist you further.

Read More
Application
CLIP
Sample
Human Cell lysate - whole cell (HEK 293T)
Specification
HEK 293T
Type
iCLIP
Irradiation Energy Level (mJ/cm2): 150
Wavelength (nm): 254
Positive control
hnRNP C antibody (sc-15386) bound protein A dynabeads, 0.1 mg/ml (antibody/beads)
Negative control
Protein A dynabeads without antibody bound

Abcam user community

Verified customer

Submitted Jun 02 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cultured Cells (Primary hippocampal neurons)
Specification
Primary hippocampal neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,25% Triton in PBS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Dec 30 2010

Application
Western blot
Sample
Rat Tissue lysate - whole (brain lysate differentially centrifuged)
Loading amount
5 µg
Specification
brain lysate differentially centrifuged
Gel Running Conditions
Reduced Denaturing (12 % SDS Gel)
Blocking step
Donkey serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 30 2010

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Primary hippocampal neurons)
Specification
Primary hippocampal neurons
Fixative
Methanol
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 30 2010

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