Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker

Target Antigen

Lamin B1 - Nuclear Envelope Marker

Primary Description

Mouse Anti-Lamin B1 [119D5-F1] - Nuclear Envelope Marker

Catalogue Number

IQ249

Quantity

50ul

Concentration

1mg/ml

Clone

119D5-F1

Host

Mouse 

Isotype

IgG1 kappa

Immunogen

Purified rat liver lamins

Myeloma/fusion partners

P3/X63.Ag8.653 mouse myeloma cells with spleen cells from a BALB/c

Specificity

Reacts with an epitope located C-terminal of residue 231 in lamin B1, does not cross react with lamin B2. Lamins do not appear to be universally distributed among different cell and tissue types. ab8982 has been shown to react with HeLa and 3T3 cells in immunocytochemistry. Other cell/tissue types have not been tested

Species Reactivity

Mouse, Rat, Sheep, Rabbit, Cow, Dog, Human. Does not react with Zebrafish

Format

Purified antibody in PBS containing 0.09% sodium azide;

Applications

WB, Dot Blot, ELISA, Flow Cyt, ICC/IF, IHC

Dilutions

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

Flow Cyt: 1/100 - 1/200. 

ICC: 1/20 - 1/50. 

WB: 1/100 - 1/1000. Predicted molecular weight: 67 kDa. (see Machiels et al reference and recommended protocol).

References

Degradation of nuclear matrix and DNA cleavage in apoptotic thymocytes Weaver VM ( J Cell Sci,Jan 1996) PUBMED 

Ehrnhoefer DE  et al. A quantitative method for the specific assessment of caspase-6 activity in cell culture. PLoS One 6:e27680 (2011). WB ; Mouse PUBMED

Ciciarello M  et al. Nuclear reformation after mitosis requires downregulation of the Ran GTPase effector RanBP1 in mammalian cells. Chromosoma 119:651-68 (2010). ICC/IF ; Mouse PUBMED

Kanninen K  et al. Intrahippocampal injection of a lentiviral vector expressing Nrf2 improves spatial learning in a mouse model of Alzheimer's disease. Proc Natl Acad Sci U S A 106:16505-10 (2009) PUBMED

Hui RC  et al. The forkhead transcription factor FOXO3a increases phosphoinositide-3 kinase/Akt activity in drug-resistant leukemic cells through induction of PIK3CA expression. Mol Cell Biol 28:5886-98 (2008). WB ; Human PUBMED

Thompson J & Winoto A During negative selection, Nur77 family proteins translocate to mitochondria where they associate with Bcl-2 and expose its proapoptotic BH3 domain. J Exp Med 205:1029-36 (2008). WB ; Mouse PUBMED

Hui RC  et al. Doxorubicin activates FOXO3a to induce the expression of multidrug resistance gene ABCB1 (MDR1) in K562 leukemic cells. Mol Cancer Ther 7:670-8 (2008). WB ; Human PUBMED

Sharma S & Brosh RM Human RECQ1 is a DNA damage responsive protein required for genotoxic stress resistance and suppression of sister chromatid exchanges. PLoS ONE 2:e1297 (2007). WB ; Human PUBMED

Ivorra C  et al. A mechanism of AP-1 suppression through interaction of c-Fos with lamin A/C. Genes Dev 20:307-20 (2006) PUBMED

Johnson MD  et al. Proteome analysis of DNA damage-induced neuronal death using high throughput mass spectrometry. J Biol Chem 279:26685-97 (2004) PUBMED

Machiels BM  et al. Nuclear lamin expression in normal testis and testicular germ cell tumours of adolescents and adults. J Pathol 182:197-204 (1997) PUBMED

Database links

Entrez Gene: 540643 Cow

Entrez Gene: 474663 Dog

Entrez Gene: 4001 Human

Entrez Gene: 16906 Mouse

Entrez Gene: 116685 Rat

Omim: 150340 Human

SwissProt: P20700 Human

SwissProt: P14733 Mouse

SwissProt: P70615 Rat

Unigene: 89497 Human

Unigene: 4105 Mouse

Unigene: 11362 Rat

Research areas

Apoptosis; Cell Biology; Class V Lamins; Cytoskeleton; Cytoskeleton / ECM; Intermediate Filaments; Lamins; Nuclear Envelope; 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 B1 antibody [119D5-F1] - Nuclear Envelope Marker

An important part of the 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 alpha helices and a carboxy terminal 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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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.


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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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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 Class V Lamins Cytoskeleton Cytoskeleton / ECM Intermediate Filaments Lamins Nuclear Envelope 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 Comparison of A and B-type lamin expression in reactive lymph nodes and nodular sclerosing Hodgkins disease - Jansen MP Degradation of nuclear matrix and DNA cleavage in apoptotic thymocytes - Weaver VM Distinct nuclear assembly pathways for lamins A and C lead to their increase during quiescence in Swiss 3T3 cells - Pugh G E Lamin A is part of the internal nucleoskeleton of human erythroleukemia cells - Neri LM Nuclear lamin expression in normal testis and testicular germ cell tumours of adolescents and adults - Machiels B M

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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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