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hnRNP A1 Antibody [9H10]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-hnRNP A1 antibody [9H10] staining human lung. Staining is localized to the nucleus.

Catalogue Number: IQ206

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Target Antigen

hnRNP A1 

Primary Description

Mouse Anti-hnRNP A1 [9H10]

Catalogue Number

IQ206

Quantity

0.1ml

Concentration

1mg/ml

Clone

9H10

Host

Mouse

Isotype

 IgG2b

Immunogen

Full length hnRNP-A1 native protein (partially purified) from HeLa cells (human)

Myeloma/fusion partners

Balb/c Sp2/0 cell line

Species Reactivity

Mouse, Human

Purification

Protein A

Format

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

Applications

Flow Cyt, IHC-P, ELISA, WB, IP, ICC/IF

Dilutions

Optimal antibody dilution should be determined by titration, however as a guideline try;

Flow Cyt: Use 1µg for 106 cells.

IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol

WB: Use at 1:2000-1:4000. Detects a band of approximately 34 kDa (predicted molecular weight: 38 kDa)

IP: Use at an assay dependent dilution. This antibody does not IP the hnRNP complex

References

RNA binding specificity of hnRNP A1: significance of hnRNP A1 highaffinity binding sites in premRNA splicing. Burd CG ( Embo J,Mar 1994) PUBMED 

Kashima T et al. hnRNP A1 functions with specificity in repression of SMN2 exon 7 splicing. Hum Mol Genet. 2007 Dec 15;16(24):3149-59. Epub 2007 Sep 19. WB, IP ; Human PMID: 17884807

Woolley AG  et al. Prognostic Association of YB-1 Expression in Breast Cancers: A Matter of Antibody. PLoS One 6:e20603 (2011). ICC/IF ; Human PUBMED

Kocgozlu L  et al. Selective and uncoupled role of substrate elasticity in the regulation of replication and transcription in epithelial cells. J Cell Sci 123:29-39 (2010). ICC/IF ; Marsupials PUBMED

Rosenberger S  et al. Alternative splicing of human papillomavirus type-16 E6/E6* early mRNA is coupled to EGF signaling via Erk1/2 activation. Proc Natl Acad Sci U S A : (2010). WB ; Human PUBMED

Sellier C  et al. Sam68 sequestration and partial loss of function are associated with splicing alterations in FXTAS patients. EMBO J 29:1248-61 (2010) PUBMED

Read EK & Digard P Individual influenza A virus mRNAs show differential dependence on cellular NXF1/TAP for their nuclear export. J Gen Virol 91:1290-301 (2010). WB, In Situ Hybridisation ; Human PUBMED

Beauchamp P  et al. The cleavage of HuR interferes with its transportin-2-mediated nuclear import and promotes muscle fiber formation. Cell Death Differ 17:1588-99 (2010). WB ; Mouse PUBMED

Lieberthal JG  et al. The role of YY1 in reduced HP1alpha gene expression in invasive human breast cancer cells. Breast Cancer Res 11:R42 (2009). WB ; Human PUBMED

van der Giessen K & Gallouzi IE Involvement of transportin 2-mediated HuR import in muscle cell differentiation. Mol Biol Cell 18:2619-29 (2007). WB, ICC/IF ; Mouse PUBMED

Database links

Clone ID: BC033714 Human

Entrez Gene: 3178 Human

Entrez Gene: 15382 Mouse

Omim: 164017 Human

SwissProt: P09651 Human

SwissProt: P49312 Mouse

Unigene: 546261 Human

Unigene: 655424 Human

Unigene: 237064 Mouse

Unigene: 432031 Mouse

Research areas

RNA binding; RNA Processing;

Also known as

HNRNPA 1 antibody;Helix destabilizing protein antibody;Helix-destabilizing proteinantibody;Heterogeneous nuclear ribonucleoprotein A1 antibody;Heterogeneous nuclear ribonucleoprotein A1B proteinantibody;Heterogeneous nuclear ribonucleoprotein B2 protein antibody;Heterogeneous nuclear ribonucleoprotein coreprotein A1 antibody;hnRNP A1 antibody;hnRNP core protein A1 antibody;HNRNPA1 antibody;HNRPA1antibody;MGC102835 antibody;Nuclear ribonucleoprotein particle A1 protein antibody;ROA1_HUMANantibody


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

Print the Datasheet for hnRNP A1 Antibody [9H10]

  • This protein belongs to the A/B subfamily of ubiquitously expressed heterogeneous nuclear ribonucleoproteins (hnRNPs) The hnRNPs are RNA binding proteins and they complex with heterogeneous nuclear RNA (hnRNA) These proteins are associated with pre mRNAs in the nucleus and appear to influence pre mRNA processing and other aspects of mRNA metabolism and transport While all of the hnRNPs are present in the nucleus some seem to shuttle between the nucleus and the cytoplasm The hnRNP proteins have distinct nucleic acid binding properties hnRNP A1 has two repeats of quasi RRM domains that bind to RNAs It is one of the most abundant core proteins of hnRNP complexes and it is localized to the nucleoplasm This protein along with other hnRNP proteins is exported from the nucleus probably bound to mRNA and is immediately re imported Its M9 domain acts as both a nuclear localization and nuclear export signal The encoded protein is involved in the packaging of pre mRNA into hnRNP particles transport of poly A mRNA from the nucleus to the cytoplasm and may modulate splice site selection It is also thought to have a primary role in the formation of specific myometrial protein species in parturition Multiple alternatively spliced transcript variants have been found for the hnRNP A1 gene but only two transcripts are fully described These variant have multiple alternative transcription initiation sites and multiple polyA sites

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.

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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
RNA binding RNA Processing
Research
hnRNP A1 functions with specificity in repression of SMN2 exon 7 splicing - Tsuyoshi Kashima RNA binding specificity of hnRNP A1 significance of hnRNP A1 high-affinity binding sites in pre-mRNA splicing - Burd CG

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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