Phospho-Lamin A/C (Ser301) Antibody - #AF7177

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Product: Phospho-Lamin A/C (Ser301) Antibody
Catalog: AF7177
Description: Rabbit polyclonal antibody to Phospho-Lamin A/C (Ser301)
Application: WB IF/ICC
Cited expt.: WB, IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 74,65kDa; 74kD(Calculated).
Uniprot: P02545
RRID: AB_2843617

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(90%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-Lamin A/C (Ser301) Antibody detects endogenous levels of Lamin A/C only when phosphorylated at Ser301.
RRID:
AB_2843617
Cite Format: Affinity Biosciences Cat# AF7177, RRID:AB_2843617.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

70 kDa lamin; Cardiomyopathy dilated 1A (autosomal dominant); CDCD1; CDDC; CMD1A; CMT2B1; EMD2; FPL; FPLD; FPLD2; HGPS; IDC; Lamin A; Lamin A/C; Lamin A/C like 1; Lamin; Lamin C; Lamin-A/C; LDP1; LFP; LGMD1B; Limb girdle muscular dystrophy 1B (autosomal dominant); LMN 1; LMN A; LMN C; LMN1; LMNA; LMNA_HUMAN; LMNC; LMNL1; Prelamin A/C; PRO1; Renal carcinoma antigen NY REN 32; Renal carcinoma antigen NY-REN-32; Renal carcinoma antigen NYREN32;

Immunogens

Immunogen:

A synthesized peptide derived from human Lamin A/C around the phosphorylation site of Ser301.

Uniprot:

P02545(LMNA_HUMAN) >>Visit HPA database.

Gene(ID):
LMNA(4000)
Expression:
P02545 LMNA_HUMAN:

In the arteries, prelamin-A/C accumulation is not observed in young healthy vessels but is prevalent in medial vascular smooth muscle cells (VSMCs) from aged individuals and in atherosclerotic lesions, where it often colocalizes with senescent and degenerate VSMCs. Prelamin-A/C expression increases with age and disease. In normal aging, the accumulation of prelamin-A/C is caused in part by the down-regulation of ZMPSTE24/FACE1 in response to oxidative stress.

Sequence:
METPSQRRATRSGAQASSTPLSPTRITRLQEKEDLQELNDRLAVYIDRVRSLETENAGLRLRITESEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVREEFKELKARNTKKEGDLIAAQARLKDLEALLNSKEAALSTALSEKRTLEGELHDLRGQVAKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETRLVEIDNGKQREFESRLADALQELRAQHEDQVEQYKKELEKTYSAKLDNARQSAERNSNLVGAAHEELQQSRIRIDSLSAQLSQLQKQLAAKEAKLRDLEDSLARERDTSRRLLAEKEREMAEMRARMQQQLDEYQELLDIKLALDMEIHAYRKLLEGEEERLRLSPSPTSQRSRGRASSHSSQTQGGGSVTKKRKLESTESRSSFSQHARTSGRVAVEEVDEEGKFVRLRNKSNEDQSMGNWQIKRQNGDDPLLTYRFPPKFTLKAGQVVTIWAAGAGATHSPPTDLVWKAQNTWGCGNSLRTALINSTGEEVAMRKLVRSVTVVEDDEDEDGDDLLHHHHGSHCSSSGDPAEYNLRSRTVLCGTCGQPADKASASGSGAQVGGPISSGSSASSVTVTRSYRSVGGSGGGSFGDNLVTRSYLLGNSSPRTQSPQNCSIM

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Dog
100
Xenopus
100
Rabbit
100
Chicken
90
Sheep
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

Function:

Lamins are components of the nuclear lamina, a fibrous layer on the nucleoplasmic side of the inner nuclear membrane, which is thought to provide a framework for the nuclear envelope and may also interact with chromatin. Lamin A and C are present in equal amounts in the lamina of mammals. Plays an important role in nuclear assembly, chromatin organization, nuclear membrane and telomere dynamics. Required for normal development of peripheral nervous system and skeletal muscle and for muscle satellite cell proliferation. Required for osteoblastogenesis and bone formation. Also prevents fat infiltration of muscle and bone marrow, helping to maintain the volume and strength of skeletal muscle and bone. Required for cardiac homeostasis.

Prelamin-A/C can accelerate smooth muscle cell senescence. It acts to disrupt mitosis and induce DNA damage in vascular smooth muscle cells (VSMCs), leading to mitotic failure, genomic instability, and premature senescence.

PTMs:

Increased phosphorylation of the lamins occurs before envelope disintegration and probably plays a role in regulating lamin associations.

Proteolytic cleavage of the C-terminal of 18 residues of prelamin-A/C results in the production of lamin-A/C. The prelamin-A/C maturation pathway includes farnesylation of CAAX motif, ZMPSTE24/FACE1 mediated cleavage of the last three amino acids, methylation of the C-terminal cysteine and endoproteolytic removal of the last 15 C-terminal amino acids. Proteolytic cleavage requires prior farnesylation and methylation, and absence of these blocks cleavage.

Sumoylation is necessary for the localization to the nuclear envelope.

Farnesylation of prelamin-A/C facilitates nuclear envelope targeting.

Subcellular Location:

Nucleus. Nucleus envelope. Nucleus lamina. Nucleus>Nucleoplasm.
Note: Farnesylation of prelamin-A/C facilitates nuclear envelope targeting and subsequent cleavage by ZMPSTE24/FACE1 to remove the farnesyl group produces mature lamin-A/C, which can then be inserted into the nuclear lamina. EMD is required for proper localization of non-farnesylated prelamin-A/C.

Nucleus speckle.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

In the arteries, prelamin-A/C accumulation is not observed in young healthy vessels but is prevalent in medial vascular smooth muscle cells (VSMCs) from aged individuals and in atherosclerotic lesions, where it often colocalizes with senescent and degenerate VSMCs. Prelamin-A/C expression increases with age and disease. In normal aging, the accumulation of prelamin-A/C is caused in part by the down-regulation of ZMPSTE24/FACE1 in response to oxidative stress.

Family&Domains:

Belongs to the intermediate filament family.

Research Fields

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Human Diseases > Cardiovascular diseases > Hypertrophic cardiomyopathy (HCM).

· Human Diseases > Cardiovascular diseases > Arrhythmogenic right ventricular cardiomyopathy (ARVC).

· Human Diseases > Cardiovascular diseases > Dilated cardiomyopathy (DCM).

References

1). Nuclear lamin A/C phosphorylation by loss of androgen receptor leads to cancer-associated fibroblast activation. Nature communications, 2024 (PubMed: 39266569) [IF=16.6]

Application: WB    Species: human    Sample:

Fig. 4: Loss of AR compromises the association of lamin A/C with the phosphatase PPP1 and results in increased lamin A/C phosphorylation at Ser 301. A Cytoscape network of proteins with reduced association with lamin A/C in AR-silenced HDFs. Proteins were identified by LC-MS/MS analysis from three different HDF strains with/without AR silencing. The list of interactors is shown in Supplementary Data S1. Names of proteins with reduced association (log fold change >1.5) in at least two strains are displayed. The inset shows PPP1 subunits associated with lamin A/C in an AR-dependent manner. B Co-immunoprecipitation assays with HDFs plus/minus AR silencing with anti-lamin A/C antibodies or nonimmune IgGs followed by immunoblotting with anti-AR, -lamin A/C, and -PPP1CA/B antibodies, with parallel analysis of the inputs. C PLA with anti-lamin A/C and PPP1CA/B antibodies of HDFs plus/minus AR silencing. Shown are representative images and quantification of the number of puncta per cell, with the mean value as red bar. Non-parametric one-way ANOVA, n = 53 cells per condition. Scale: 10 µm. D Immunoblot analysis of HDFs treated with ARCC4 (1 µM) or AZD3514 (10 µM) for 72 h, with phospho-Ser301 lamin A/C, total lamin A/C, and β-Actin antibodies. E Immunofluorescence analysis of three HDF strains treated with ARCC4 (1 µM) or EtOH for 72 h, antibodies against phospho-Ser301-lamin A/C (yellow), and DAPI for nuclear staining. Representative images (left) and fluorescence signal intensity quantification (right), Two-tailed unpaired t-test, n = 3 strains (50 cells per condition), mean ± SE. Scale: 10 µm. F IF analysis of three HDF strains, infected with AR-silencing lentiviruses versus control, with antibodies against phospho-Ser301(pSer301)-lamin A/C (yellow) and counterstained with DAPI. Representative images (left) and fluorescence intensity quantification (right), nonparametric one-way ANOVA, n = 3 strains (50 cells per condition), mean ± SE. Scale: 10 µm. Fluorescence intensities per individual counted cells for each strain are shown in Supplementary Fig. 4D. G 3D Surface Reconstruction: Whole nuclei of HDFs plus/minus AR silencing stained with anti-phospho-S301-lamin A/C (red) and total lamin A/C (yellow). Z-stacks of confocal images used for 3D volumetric reconstruction with Imaris Surface tool. 2D and 3D images of Lamin A/C and pSer301 signals; top and lateral views. Scale: 5 µm. Source data for individual graphs are in the Source Data file.
2). Nuclear lamin A/C phosphorylation by loss of Androgen Receptor is a global determinant of cancer-associated fibroblast activation. bioRxiv : the preprint server for biology, 2023 (PubMed: 37425957)

Application: IF/ICC    Species: human    Sample:

Figure 5. Lamin A/C phosphorylation at Ser 301 is a signature of CAFs. A) IF analysis of CAFs and matched normal fibroblasts (HDFs) from the same patient with images of pSer301 lamin A/C (pSer301; red) and total lamin A/C (yellow). Shown are representative images and quantification of phospho-ser301-lamin A/C fluorescence signal intensity assessed by Image J and expressed in arbitrary units (AU). n(cells)>50 per condition together with mean and statistical significance, ****p50 per condition together with mean and statistical significance, ****p20 per together with mean and statistical significance, ****p50 per condition together with mean and statistical significance, ****p

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