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Gemin 3 Antibody [12H12]

Immunocytochemistry/ Immunofluorescence - Anti-Gemin 3 antibody [12H12] staining Gemin 3 in the Human HEK cell line

Catalogue Number: IQ204

£226.00
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Target Antigen

Gemin 3

Primary Description

Mouse Anti-Gemin 3 [12H12]

Catalogue Number

IQ204

Quantity

0.1ml

Concentration

 1mg/ml

Clone

12H12

Host

Mouse

Isotype

 IgG1

Immunogen

Recombinant 6His-tag COOH-terminal domain of Gemin3 (amino acids 368-548)

Myeloma/fusion partners

SP2/0;

Species Reactivity

Human, Rat, Mouse, Hamster

Purification

Protein A

Format

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

Applications

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

Dilutions

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

WB: Use at an assay dependent dilution. Detects a band of approximately 105 kDa (predicted molecular weight: 99 kDa).

Flow Cyt: Use 1µg for 106 cells.

ICC/IF: 1/50.

References

Mahmoudi S  et al. WRAP53 is essential for Cajal body formation and for targeting the survival of motor neuron complex to Cajal bodies. PLoS Biol 8:e1000521 (2010). WB ; Human PUBMED

Bradrick SS & Gromeier M Identification of gemin5 as a novel 7-methylguanosine cap-binding protein. PLoS One 4:e7030 (2009). WB ; Human PUBMED

Castelló A  et al. RNA nuclear export is blocked by poliovirus 2A protease and is concomitant with nucleoporin cleavage. J Cell Sci 122:3799-809 (2009). WB ; Human PUBMED

Walker MP  et al. SMN complex localizes to the sarcomeric Z-disc and is a proteolytic target of calpain. Hum Mol Genet 17:3399-410 (2008). ICC/IF ; Hamster PUBMED

Morency E  et al. A novel cell response triggered by interphase centromere structural instability. J Cell Biol 177:757-68 (2007). ICC/IF ; Human PUBMED

Murashov AK et al. RNAi pathway is functional in peripheral nerve axons. FASEB J. 2007 Mar;21(3):656-70. Epub 2007 Jan 5. ICC/IF, IHC, WB ; Rat Mouse PMID: 17209129

Hamamoto S et al. Identification of a novel human immunodeficiency virus type 1 integrase interactor, Gemin2, that facilitates efficient viral cDNA synthesis in vivo. J Virol. 2006 Jun;80(12):5670-7. WB ; Human PMID: 16731905

Zhang H  et al. Multiprotein complexes of the survival of motor neuron protein SMN with Gemins traffic to neuronal processes and growth cones of motor neurons. J Neurosci 26:8622-32 (2006) PUBMED

Gemin3: A novel DEAD box protein that interacts with SMN, the spinal muscular atrophy gene product, and is a component of gems. Charroux et.al ( J Cell Biol,Dec 1999) PUBMED

Database links

Entrez Gene: 11218 Human

Omim: 606168 Human

SwissProt: Q9UHI6 Human

Unigene: 591405 Human

Research areas

Neuroscience; signal transduction; Splicing;

Also known as

Component of gems 3 antibody;DDX 20 antibody;DDX20 antibody;DDX20_HUMAN antibody;DEAD (Asp Glu AlaAsp) box polypeptide 20 antibody;DEAD (Asp-Glu-Ala-Asp) box polypeptide 20 antibody;DEAD box protein 20antibody;DEAD box protein DP 103 antibody;DEAD box protein DP103 antibody;DEAD-box protein DP103antibody;DEAD/H (Asp Glu Ala Asp/His) box polypeptide 20 103kD antibody;DEAD/H (Asp Glu Ala Asp/His) boxpolypeptide 20 antibody;DEAD/H BOX 20 antibody;DKFZP434H052 antibody;DP 103 antibody;DP103 antibody;Gemin-3antibody;Gemin3 antibody;Probable ATP dependent RNA helicase DDX20 antibody;Probable ATP-dependent RNA helicaseDDX20 antibody;SMN interacting protein antibody


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

Print the Datasheet for Gemin 3 Antibody [12H12]

  • The survival of motor neurons (SMN) gene is the disease gene of spinal muscular atrophy (SMA) a common motor neuron degenerative disease The SMN protein is part of a complex containing several proteins of which one SIP1 (SMN interacting protein 1) has been characterized so far The SMN complex is found in both the cytoplasm and in the nucleus where it is concentrated in bodies called gems In the cytoplasm SMN and SIP1 interact with the Sm core proteins of spliceosomal small nuclear ribonucleoproteins (snRNPs) and they play a critical role in snRNP assembly In the nucleus SMN is required for pre-mRNA splicing likely by serving in the regeneration of snRNPs A DEAD box putative RNA helicase named Gemin 3 which is another component of the SMN complex has been identified Gemin 3 interacts directly with SMN as well as with SmB SmD2 and SmD3 Immunolocalization studies using mAbs to Gemin 3 show that it colocalizes with SMN in gems Gemin 3 binds SMN via its unique COOH-terminal domain and SMN mutations found in some SMA patients strongly reduce this interaction The presence of a DEAD box motif in Gemin 3 suggests that it may provide the catalytic activity that plays a critical role in the function of the SMN complex on RNPs

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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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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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
Neuroscience signal transduction Splicing
Research
A novel DEAD box protein that interacts with SMN the spinal muscular atrophy gene product and is a component of gems - Charroux etal Identification of a Novel Human Immunodeficiency Virus Type 1 Integrase Interactor Gemin2 That Facilitates Efficient Viral cDNA Synthesis In Vivo - Seiji Hamamoto et al

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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