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PABP Antibody [10E10]

Immunocytochemistry/ Immunofluorescence-PABP antibody [10E10] stained HeLa cells

Catalogue Number: IQ216

£226.00
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Target Antigen

PABP

Primary Description

Mouse Anti-PABP [10E10]

Catalogue Number

IQ216

Quantity

0.1ml 

Concentration

1mg/ml

Clone

10E10

Host

Mouse 

Isotype

IgG2b

Immunogen

Recombinant PABP (Human) expressed from its 1.85 kbp cDNA, NcoI to SspI

Myeloma/fusion partners

SP2/0

Species Reactivity

Rabbit, Chicken, Human, Xenopus laevis, Hamster

Purification

Protein A

Format

Purified antibody (from supernatant) containing PBS 0.1% sodium azide;

Applications

ELISA, WB, IP, ICC/IF, Flow Cyt

Dilutions

Optimal antibody dilution should be determined by titration, however try:

WB: Use at an assay dependent dilution. Detects a band of approximately 70 kDa (predicted molecular weight: 71 kDa)

ICC/IF: Use at an assay dependent dilution. PubMed: 17977970

Flow Cyt: Use 1µg for 106 cells

References

Briggs JW  et al. The ezrin metastatic phenotype is associated with the initiation of protein translation. Neoplasia 14:297-310 (2012) PUBMED

Gorgoni B  et al. Poly(A)-binding proteins are functionally distinct and have essential roles during vertebrate development. Proc Natl Acad Sci U S A 108:7844-9 (2011) PUBMED

Rattenbacher B et al. Analysis of CUGBP1 targets identifies GU-repeat sequences that mediate rapid mRNA decay. Mol Cell Biol. 2010 Aug;30(16):3970-80. doi: 10.1128/MCB.00624-10. Epub 2010 Jun 14. IP ; Human PMID: 20547756

Unsworth H  et al. mRNA escape from stress granule sequestration is dictated by localization to the endoplasmic reticulum. FASEB J 24:3370-80 (2010) PUBMED

Ogawa S et al. Physical and functional interactions between STAT5 and Runx transcription factors. J Biochem. 2008 May;143(5):695-709. doi: 10.1093/jb/mvn022. Epub 2008 Feb 22. WB ; Hamster  ICC/IF ; Human PMID: 18296717

Kanno T et al. Expression of Kaposi's sarcoma-associated herpesvirus-encoded K10/10.1 protein in tissues and its interaction with poly(A)-binding protein. Virology. 2006 Aug 15;352(1):100-9. Epub 2006 May 22. ICC/IF IP WB ; Human PMID: 16716377 

Nagaoka K et al. Stability of casein mRNA is ensured by structural interactions between the 3'-untranslated region and poly(A) tail via the HuR and poly(A)-binding protein complex. Biochim Biophys Acta. 2006 Mar-Apr;1759(3-4):132-40. Epub 2006 Apr 20. WB ; Rabbit PMID: 16714065 

Hofmann I et al. Identification of the junctional plaque protein plakophilin 3 in cytoplasmic particles containing RNA-binding proteins and the recruitment of plakophilins 1 and 3 to stress granules. Mol Biol Cell. 2006 Mar;17(3):1388-98. Epub 2006 Jan 11. ICC/IF WB IP ; Human PMID: 16407409 

Baez MV et al. Mammalian Smaug is a translational repressor that forms cytoplasmic foci similar to stress granules. J Biol Chem. 2005 Dec 30;280(52):43131-40. Epub 2005 Oct 12. ICC/IF ; Hamster PMID: 16221671

The mRNA poly(A)binding protein: localization, abundance, and RNAbinding specificity. Grlach M ( Exp Cell Res.,April 1994) PUBMED 

Database links

Entrez Gene: 26986 Human

Omim: 604679 Human

SwissProt: P11940 Human

Unigene: 387804 Human

Research areas

Chromatin; Dicer; P bo

Also known as

PAB 1 antibody;PAB1 antibody;PABP 1 antibody;PABP1 antibody;PABPC 1 antibody;PABPC1antibody;PABPC2 antibody;PABPL1 antibody;Poly A binding protein 1 antibody;Poly A binding protein cytoplasmic1 antibody;Polyadenylate binding protein 1 antibody


THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR THERAPEUTIC OR DIAGNOSTIC USE.

Print the Datasheet for PABP Antibody [10E10]

  • The poly(A)-binding protein (PABP) which is found complexed to the 3-prime poly(A) tail of eukaryotic mRNA is required for poly(A) shortening and translation initiation Grange et al (1987) isolated a melanoma cell cDNA encoding human PABP The predicted 633-amino acid protein contains 4 repeats of an approximately 80-amino acid unit in its N-terminal half The authors found that this repeat region is highly conserved between human and yeast PABP and is sufficient for poly(A) binding In vitro translation of the human PABP cDNA yielded a protein with an apparent molecular mass of 73 kD by SDS-PAGE Northern blot analysis indicated that PABP is expressed as a 29-kb mRNA in human melanoma cells Gorlach et al (1994) noted that each of the 4 repeats of PABP is a ribonucleoprotein (RNP) consensus sequence RNA-binding domain They determined that PABP has a pI of approximately 103 and is a very abundant stable protein Immunofluorescence studies of mammalian cells indicated that PABP is located exclusively in the cytoplasm However using both indirect immunofluorescence and tagging of PABP1 by fusion to the green fluorescent protein (GFP) Afonina et al (1998) demonstrated that PABP1 shuttles between the nucleus and cytoplasm PABP1 accumulated in the nucleus when transcription was inhibited suggesting that active transcription is required for nuclear export of PABP1

Immunofluorescence protocol - Formaldehyde fixation

   1. Collect cells from T.c.unit and remove media from petri dish using suction.
   2. Wash with 1x PBS and remove.
   3. Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperature on an orbital shaker.
   4. Remove PFA and incubate in 0.5% Triton X-IOO in 1x PBS for 5 minutes at room temperature.
   5. Prepare blocking reagent, this is also the antibody diluent.
   6. Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker.
   7. Block with 1 % NCS and 1x PBS for 30 minutes at room temperature.
   8. Prepare primary antibodies (50?l/coverslip) and moist staining chambers.
   9. Wash cells 2x with lx PBS at room temperature and air dry briefly.
  10. Incubate with primary antibody for 1 hr at room temperature in the dark in staining chambers. During this time prepare the secondary antibody.
  11. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
  12. Incubate with secondary antibody for 1 hour at room temperature in the dark in staining chambers.
  13. Wash cells 5x with 1x PBS.
  14. Mount in Dapi.

Solutions (prepare fresh the same day of staining).

    * 1x Phosphate buffered saline.
    * Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS).
    * Fixation solution: 3.5% Para formaldehyde.

1.75g PFA in 20 ml d.H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C until dissolved. Add 4 drops IN HCI and check pH indicator strip. PH should be 7.4. Complete volume with d.H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips.


Immunofluorescence protocol - Methanol/acetone fixation

   1. Collect cells from T.C.unit and remove media from petri dish using suction.
   2. Wash with 1x PBS and remove.
   3. Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice.
   4. Prepare blocking reagent, this is also the diluent for the antibodies.
   5. Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker.
   6. Block with 1% NCS and Ix PBS for 30 minutes at RT.
   7. Prepare primary antibodies (50?l/coverslip) and moist staining chambers.
   8. Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes.
   9. Incubate with primary antibody for 1 hr at RT in the dark in staining chambers.  During this time prepare secondary antibody.
  10. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
  11. Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers.
  12. Wash cells 5x with 1x PBS.
  13. Mount in Dapi.

Solutions (prepare fresh the same day of staining)

    * 1x Phosphate buffered saline.
    * Blocking reagent: 1% NCS in 1x PBS (use fresh 10x PBS).
    * Fixation solution: methanol:acetone 1: 1 ice cold.



Western Blotting Protocol

   1. Transfer gel to PDVF or nitrocellulose membrane
   2. Place membrane in plastic tray in blocking buffer for one hour with agitation
   3. Rinse in wash buffer
   4. Incubate in wash buffer plus primary antibody for one hour
   5. Wash 6 X 5 minutes with wash buffer
   6. Incubate in wash buffer plus secondary antibody for one hour
   7. Wash 6X 5 minutes with wash buffer
   8. Detect (e.g. ECL, Amersham according to manufacturers instructions)

Wash buffer
PBS + 0.1% Tween 20

Blocking buffer
Wash buffer + 5% dried milk powder

The concentration of antibodies used depends on each antibody, the amount of antigen and the detection method used. Generally, dilution is in the range of a few hundred times dilution to a few thousand times dilution, but usually has to be determined empirically.

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How do I place an order?
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How do I place an order?

When placing an order we require a purchase order number, plus name and contact details of the purchaser, and the person who will be using the product. We will add VAT for every order received from an EU country, unless you are VAT exempt. For such VAT exempt customers we will need to receive a copy of the exemption certificate with the order.

Orders can be placed either by our website, via email or by mail. Please see our contact us page for more details

All orders are subject to availability. Prices of products do not include shipping, VAT or import duties, where these are applicable.
Price and other information provided are subject to change without notice. While every effort is made to keep information provided Up to date, ImmuQuest will not be liable if errors should occur in such information.

Formal acceptance of an order will take place when the goods are dispatched. If prices should be changed between the time of receipt of an order and dispatch, ImmuQuest will contact you in advance.


Ordering via the website

To order via the website you will need to find the product(s) you are looking for by either using our search, browsing our categories or using our full antibody catalogue.

Once you have found the antibodies you are looking for you can view more detailed information about a specific antibody by clicking the product or the 'View' button. Once you are sure it is the antibody you are looking for you can add it to your order by clicking the 'Add To Order' button.

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How long does delivery take?

Delivery usually takes no more than one week from reciept of an order.

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Do you guarantee your antibodies?

Yes. Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.

If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.

Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.

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Do you charge VAT?

If you are in the EU and you are no exempt from paying VAT then we must charge VAT (17.5%).

Customers that are exempt need to fax an exemption form showing their vat exemption number.

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You can still place an order without providing a purchase order number but we must recieve payment before your order is despatched.

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Non-website orders

Non-website orders can be placed directly with Immuquest at sales@immuquest.com or click on the "contact us" tab for address details

 

Product Guarantee

We are strongly committed to providing the best quality products, and the best service possible. In order to do this we depend on your feedback.

Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.

If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.

Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.

If you have any questions please contact us using the following details:
ImmuQuest Ltd
26 The Green
Seamer
North Yorkshire
TS9 5LP
UK

Tel: 01642 713533
Fax: 01642 713988
International Tel: +44 1642 713533
International Fax: +44 1642 713988

When placing an order ImmuQuest require a purchase order number, plus name and contact details of the purchaser, and the person who will be using the product. ImmuQuest will also need a VAT number for customers in the European Union.UK customers that are VAT exempt need to fax an exemption certificate.

Orders can be placed either by our website, via email or by mail. Please see our contact us page for more details. All orders are subject to availability. Prices of products do not include shipping, VAT or import duties where these are applicable.

Price and other information provided are subject to change without notice. While every effort is made to keep information provided Up to date, ImmuQuest will not be liable if errors should occur in such information. Formal acceptance of an order will take place when the goods are dispatched. If prices should be changed between the time of receipt of an order and dispatch, ImmuQuest will contact you in advance.

Research Areas
Chromatin Dicer P bodies/GW bodies Regulation RNAi
Research
Expression of Kaposis sarcoma-associated herpesvirus-encoded K10/101 protein in tissues and its interaction with poly(A)-binding protein - Takayuki Kanno et al Identification of the Junctional Plaque Protein Plakophilin 3 in Cytoplasmic Particles Containing RNA-binding Proteins and the Recruitment of Plakophilins 1 and 3 to Stress Granules - Ilse Hofmann et al Mammalian Smaug Is a Translational Repressor That Forms Cytoplasmic Foci Similar to Stress Granules - Mara V Baez et al The mRNA poly(A)-binding protein localization abundance and RNA-binding specificity - Grlach M

Offline Orders

For offline orders please call +44 (0) 1642 713533 or click here to email us.

Delivery & returns

We use federal Express for UK & international shipments, andfor standard shipments the cost is £30 UK & £55 international. If the customer uses their account, we charge a £20 handling fee (UK & international)

We are strongly committed to providing the best quality products, and the best service possible.

Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.

If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.

Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.

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