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BM4503  Lamin B (B1+B2) antibody

see related secondary antibodies
see all 3 Lamin B products
2 ml / US$ 525
NOVUS BIOLOGICALS
PO Box 802 Littleton, CO 80160, USA
E-Mail: novus@novus-biologicals.com
Homepage: http://www.novus-biologicals.com

Quick Overview

Mouse anti Lamin B (B1+B2) X223

Synonyms

filament-type protein,

Product Description

Mouse anti Lamin B (B1+B2) X223, Presentation: Supernatant. Product is tested for Immunofluorescence ( IF ), Western blot / Immunoblot ( WB )

Properties

ApplicationsImmunofluorescence ( IF ), Western blot / Immunoblot ( WB )
ReactivityHuman ( Hu ), Bovine ( Bov ), Rat ( Rt ), Xenopus ( Xen )
PresentationSupernatant
HostMouse
IsotypeIgG1
CloneX223
Catalog NumberBM4503
Quantity2 ml
PriceUS$ 525
DeliveryWorldwide
ManufacturerAcris Antibodies GmbH
Datasheetview PDF-Download
BM4503.pdf

Datasheet Extract

Alternate namesLMN2, LMNB, LMNB2, MGC111419
ImmunogenNuclear pore complex-lamina fraction of Xenopus laevis (XLKE-A6 cells).
Format
State: Tissue culture supernatant with 0.09% sodium azide as preservative.
ApplicationsSuitable for Immunofluorescence and Western blot (1/50, ECL technique).
SpecificityThis antibody decorates the karyoskeleton, i.e. the intermediate filament equivalent of the nucleus.
Reacts with lamin isotypes of Mr 60-75 kD.
Recognises lamin B1 & B2 in human, bovine & rat, and lamin B2 in mouse.
StorageStore the antibody at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Shelf life: One year from despatch.
References1. Krohne G. and Benavente R.: The Nuclear Lamins. A Multigene Family of Proteins in Evolution and Differentiation. Exp. Cell Res. 162, 1-10 (1986)
2. Franke, W.W.: Nuclear Lamins and Cytoplasmic Intermediate Filament Proteins: A Growing Multigene Family. Cell 48, 3-4 (1987)
3. Höger, T.H., Grund, C., Franke, W.W., Krohne G.: Immunolocalization of Lamins in the thick nuclear lamina of human synovial cells.Europ J Cell Biol 54, 150-156 (1991)
4. Höger TH., Zatloukal K, Waizenegger I, and Krohne G: Characterization of a second highly conserved B-type lamin present in cells previously thought to contain only a single B-type lamin. Chromosoma 99, 379-390 (1990)
5. Alsheimer M, von Glasenapp E, Schnölzer M, Heid H and Benavente R: Meiotic lamin C: The unique amino-terminal hexapeptide GNAEGR is essential for nuclear envelope association. Proc.Natl.Acad.Sci. USA 97, 13120-13125 (2000)
ProtocolsImmunofluorescence protocol - Formaldehyde fixation
Collect cells from T.c.unit and remove media from petri dish using suction.
Wash with 1x PBS and remove.
Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperature on an orbital shaker.
Remove PFA and incubate in 0.5% Triton X-IOO in 1x PBS for 5 minutes at room temperature.
Prepare blocking reagent, this is also the antibody diluent.
Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker.
Block with 1 % NCS and 1x PBS for 30 minutes at room temperature.
Prepare primary antibodies (50μl/coverslip) and moist staining chambers.
Wash cells 2x with lx PBS at room temperature and air dry briefly.
Incubate with primary antibody for 1 hr at room temperature in the dark in staining chambers. During this time prepare the secondary antibody.
Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
Incubate with secondary antibody for 1 hour at room temperature in the dark in staining chambers.
Wash cells 5x with 1x PBS.
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
Collect cells from T.C.unit and remove media from petri dish using suction.
Wash with 1x PBS and remove.
Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice.
Prepare blocking reagent, this is also the diluent for the antibodies.
Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker.
Block with 1% NCS and Ix PBS for 30 minutes at RT.
Prepare primary antibodies (50μl/coverslip) and moist staining chambers.
Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes.
Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. During this time prepare secondary antibody.
Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers.
Wash cells 5x with 1x PBS.
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
Transfer gel to PDVF or nitrocellulose membrane
Place membrane in plastic tray in blocking buffer for one hour with agitation
Rinse in wash buffer
Incubate in wash buffer plus primary antibody for one hour
Wash 6 X 5 minutes with wash buffer
Incubate in wash buffer plus secondary antibody for one hour
Wash 6X 5 minutes with wash buffer
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.



14 Secondary Antibodies

Catalog No.Name / HostPresentationReactivity 
R1253B Mouse IgG (H&L)  
  Rabbit Biotin   2 mg / US$ 315
   
R1253F Mouse IgG (H&L)  
  Rabbit FITC   2 mg / US$ 295
   
R1253T Mouse IgG (H&L)  
  Rabbit TRITC   2 mg / US$ 295
   
R1253TR Mouse IgG (H&L)  
  Rabbit Texas Red   2 mg / US$ 305
   
R1253HRP Mouse IgG (H&L)  
  Rabbit HRP   2 mg / US$ 325
   
R1253AP Mouse IgG (H&L)  
  Rabbit AP   1 mg / US$ 335
   
R1457C2 Mouse IgG (H&L) multi-species ads.  
  Goat Cy2   1 mg / US$ 455
   
R1457C3 Mouse IgG (H&L) multi-species ads.  
  Goat Cy3   1 mg / US$ 455
   
R1457C35 Mouse IgG (H&L) multi-species ads.  
  Goat Cy3.5   1 mg / US$ 455
   
R1457C5 Mouse IgG (H&L) multi-species ads.  
  Goat Cy5   1 mg / US$ 455
   
R1457C55 Mouse IgG (H&L) multi-species ads.  
  Goat Cy5.5   1 mg / US$ 455
   
R1405R Mouse IgG (H&L) F(ab')2 Fragment ads. to Bov, Eq, Hu, Rb, Rt, Sh  
  Goat PE   1 ml / US$ 465
   
R1403R Mouse IgG (H&L) F(ab')2 Fragment ads. to Hu serum proteins  
  Rabbit PE   1 ml / US$ 465
   
R1455R Mouse IgG (H&L) F(ab')2 Fragment multi-species ads.  
  Donkey PE   1 ml / US$ 465
   

Click here to see all secondary antibodies for 'BM4503 Lamin B (B1+B2)'.