Lamin-A/C (LMNA) - BM4505S | acris-antibodies.com

Product Description for Lamin-A/C (LMNA)

Mouse anti Human, Mouse Lamin-A/C (LMNA) JOL5.
Presentation: Supernatant
Product is tested for Immunocytochemistry/Immunofluorescence, Western blot / Immunoblot.

Properties for Lamin-A/C (LMNA)

Product Category	Antibodies
Quantity	1 ml
Synonyms	70 kDa Lamin, FocusOn028, FocusOn098, LMN1, LMNA, Lamin A, Lamin A + C, Lamin-A/C, NY-REN-32, NYREN32, Nuclear Envelope Marker, Renal carcinoma antigen NY-REN-32
Presentation	Supernatant
Reactivity	Hu, Ms
Applications	ICC/IF, WB
Clonality	Monoclonal
Clone	JOL5
Host	Mouse
Isotype	IgG2
Shipping to	Worldwide
PDF datasheet	View Datasheet
Manufacturer	Acris Antibodies GmbH

Datasheet Extract

Immunogen	Swiss Prot Num: P02545 Immunogen: Recombinant bacterially expressed human lamin A. Genename: 4000
Antibody type	Ab
Application	Immunofluorescence: 1/10 (as a guideline). Western blot (ECL technique): 1/50.
Background	Nuclear Lamins form a network of intermediate-type filaments at the nucleoplasmic site of the nuclear membrane. Two main subtypes of nuclear lamins can be distinguished, i.e. 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, i.e. Lamin A, Lamin C and Lamin Adel 10, while the B-type Lamins include two proteins arising from two distinct genes, i.e. Lamin B1 and Lamin B2. Recent evidence has revealed that mutations in A-type Lamins give rise to a range of rare but dominant genetic disorders, including Emery-Dreifuss muscular dystrophy, dilated cardiomyopathy with conduction-system disease and Dunnigan-type familial partial lipodystrophy. In addition, the expression of A-type Lamins coincides with cell differentiation and as A-type Lamins specifically interact with chromatin, a role in the regulation of differential gene expression has been suggested for A-type Lamins.
Protocols	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 (50μl/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 (50μl/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
General Readings	Dyer JA, Kill IR, Pugh G, Quinlan RA, Lane EB, Hutchison CJ. Cell cycle changes in A-type lamin associations detected in human dermal fibroblasts using monoclonal antibodies. Chromosome Res. 1997 Sep;5(6):383-94. PubMed PMID: 9364940.
Storage	Store 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.
Format	Purification: Unpurified. State: Tissue Culture Supernatant with 0.09% Sodium Azide as preservative. Supernatant
Specificity	Specificity: Detects an un-phosphorylated form of lamin A/C and the epitope is mapped to the N-terminal head domain of both proteins. Species: Human and mouse.
Gene ID	4000
FocusOn and Reviews	FocusOns: Antibodies to Lamins - FocusOn 028
Targets	Lamin-A/C (LMNA) antibody

Accessory Products

Proteins and/or Positive Controls

Positive controls for Lamin-A/C (LMNA) (3 products)

Catalog No.	Species
H00004000-T01	Lamin-A/C (LMNA) Lysate(Denatured)
		0.1 ml / €240.00
		Abnova Taiwan Corp.
NBL1-12564	Lamin A Lysate
		0.1 mg / €370.00
		Novus Biologicals Inc.
NBL1-12565	Lamin A Lysate
		0.1 mg / €370.00
		Novus Biologicals Inc.

Recommended Secondary Antibodies for Lamin-A/C (LMNA) (8 products)

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R1253HRP	Mouse IgG (H+L chain) antibody
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R1253TR	Mouse IgG (H+L chain) antibody
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R1403R	Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody
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R1405R	Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody
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R1455R	Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody
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