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Lamin B2 antibody [LN43] - Nuclear Envelope Marker

Immunostaining reaction of lamin B2 antibody [LN43] with human epidermis

Catalogue Number: IQ250

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Target Antigen

Lamin B2 - Nuclear Envelope Marker

Primary Description

Mouse Anti-Lamin B2 [LN43] - Nuclear Envelope Marker

Catalogue Number

IQ250

Quantity

50ul

Concentration

1mg/ml

Clone

LN43

Host

Mouse 

Isotype

IgG1

Immunogen

Detergent insoluble fraction of potoroo cell line PtK1.

Myeloma/fusion partners

Mouse myeloma cells with spleen cells from a mouse

Specificity

Reacts with an epitope located in the C-terminal part of lamin B2.

Species Reactivity

 Mouse, Hamster, Human, Xenopus laevis, Zebrafish

Format

Purified antibody in PBS containing 0.09% sodium azide

Applications

ICC/IF, Flow Cyt, IHC-Fr, WB, IP

Dilutions

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

ICC: 1/400.

Flow Cyt: Use 1µg for 106 cells.

IHC-Fr: 1/1 - 1/1. (when used with an avidin-biotinylated HRP complex as detection agent).

WB: 1/100 - 1/1000. Detects a band of approximately 65-67 kDa.

References

Tsai YJ  et al. Ribosome distribution in HeLa cells during the cell cycle. PLoS One 7:e32820 (2012). ICC/IF ; Human PUBMED

Shimi T  et al. The role of nuclear lamin B1 in cell proliferation and senescence. Genes Dev 25:2579-93 (2011) PUBMED

Ishimoto K  et al. Sterol-mediated Regulation of Human Lipin 1 Gene Expression in Hepatoblastoma Cells. J Biol Chem 284:22195-205 (2009). WB ; Human PUBMED

Mackay DR  et al. The nucleoporin Nup153 has separable roles in both early mitotic progression and the resolution of mitosis. Mol Biol Cell 20:1652-60 (2009). WB ; Human PUBMED

Taimen P  et al. A progeria mutation reveals functions for lamin A in nuclear assembly, architecture, and chromosome organization. Proc Natl Acad Sci U S A : (2009). WB ; Human PUBMED

Hisatsune J  et al. Molecular Characterization of Helicobacter pylori VacA Induction of IL-8 in U937 Cells Reveals a Prominent Role for p38MAPK in Activating Transcription Factor-2, cAMP Response Element Binding Protein, and NF-{kappa}B Activation. J Immunol 180:5017-27 (2008). WB ; Human PUBMED

Koutsami MK  et al. Centrosome abnormalities are frequently observed in non-small-cell lung cancer and are associated with aneuploidy and cyclin E overexpression. J Pathol 209:512-21 (2006) PUBMED

Tsai MY  et al. A mitotic lamin B matrix induced by RanGTP required for spindle assembly. Science 311:1887-93 (2006). ICC/IF ; Human PUBMED

Broers JL  et al. A- and B-type lamins are differentially expressed in normal human tissues. Histochem Cell Biol 107:505-17 (1997) PUBMED

Krohne G. Lamins. Methods Cell Biol. 2004;78:573-96 WB, ICC/IF, IP. Mouse, Human PMID: 15646632`

Database links

Entrez Gene: 84823 Human

Entrez Gene: 16907 Mouse

Omim: 150341 Human

SwissProt: Q03252 Human

SwissProt: P21619 Mouse

Unigene: 728836 Human

Unigene: 7362 Mouse

Research areas

Apoptosis; Cell Biology; Chromatin; Class V Lamins; Cytoskeleton; Cytoskeleton / ECM; Intermediate Filaments; Lamins; Nucleoplasm; Nucleus; Subcellular Markers; Tags Cell Markers;

Also known as

Lamin B1 antibody;Lamin B2 antibody;LMN antibody;LMN B2 antibody;LMN2 antibody;LMNBantibody;LMNB1 antibody;LMNB2 antibody


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

Print the Datasheet for Lamin B2 antibody [LN43] - Nuclear Envelope Marker

  • An important part of the cell nucleus is formed by nuclear lamina Nuclear lamins form a network of filaments at the nucleoplasmic site of the nuclear membrane Two main subtypes of nuclear lamins can be distinguished ie A-type lamins and B-type lamins The A-type lamins comprise a set of three proteins arising from the same gene by alternative splicing ie lamin A lamin C and lamin Adel10 while the B-type lamins include two proteins arising from two distinct genes ie lamin B1 and lamin B2 The nuclear lamins comprise a unique subclass of the intermediate filament protein family They share a molecular domain organisation with the other intermediate filament proteins in that they are fibrous molecules that have an aminoterminal globular head a central rod of a-helices and a carboxyterminal globular domain Many biochemical and molecular features of lamins have been studied but their functions remain still largely undetermined One of the functions ascribed to the lamina is the maintenance of the structural integrity of the nucleus Besides interactions with the nuclear membrane and other intermediate filaments lamins interact with the nuclear chromatin Eukaryotic chromatin is organised into loops which are attached to the nuclear matrix This organisation is thought to contribute to compaction of the chromatin and regulation of gene expression Lamins as part of the nuclear matrix may be involved in these processes since chromatin binding sites have been detected in both A- and B-type lamins

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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What if I don't have a purchase order number?

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
Apoptosis Cell Biology Chromatin Class V Lamins Cytoskeleton Cytoskeleton / ECM Intermediate Filaments Lamins Nucleoplasm Nucleus Subcellular Markers Tags Cell Markers
Research
A- and B-type lamins are differentially expressed in normal human tissues - Broers JL Abnormal A-type lamin organization in a human lung carcinoma cell line - Machiels BM An alternative splicing product of the lamin A/C gene lacks exon 10 - Machiels BM Zorenc AH Endert JM Kuijpers HJ van Eys GJ Ramaekers FC Broers JL Comparison of A and B-type lamin expression in reactive lymph nodes and nodular sclerosing Hodgkins disease - Jansen MP Internal lamin structures within G1 nuclei of human dermal fibroblasts - Bridger JM Lamin proteins form an internal nucleoskeleton as well as a peripheral lamina in human cells - Hozak P Nuclear lamin expression in normal testis and testicular germ cell tumours of adolescents and adults - Machiels B M Nuclei that lack a lamina accumulate karyophilic proteins and assemble a nuclear matrix - Jenkins H Hlman T Lyon C Lane B Stick R Hutchison C

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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