Heart Neuronal crest derivative expressed 2

Alternate Nomenclature: NP_073187;

Hand proteins are evolutionary conserved basic helix-loop-helix (bHLH) transcription factors implicated in development of neural crest-derived tissues, heart and limbs. There are two isofroms of Hand protein expressed in embryonic tissue (Hand 1 and Hand 2). The basic helix-loop-helix (bHLH) transcription factors Hand1 and Hand2 are essential for embryonic development, Hand1 is expressed in the ventral zone of the bronchial arches, while the Hand2 protein is expressed ventrolaterally across two-thirds of the mandibular arch (1). HAND2 plays an essential role in cardiac morphogenesis. The prevalence of HAND2 mutations in congenial heart disease (CHD) and the correlation between the HAND2 genotype and CHD phenotype have been shown. Although highly evolutionarily conserved genes, HAND cardiac expression patterns differ across species (3).  Heart formation requires the fusion of bilateral cardiomyocyte populations as they move towards the embryonic midline. The Hand2 transcription factor is essential for cardiac fusion; however, the effector genes that execute this function of Hand2 are unknown.

HAND2 is located on chromosome 4q33 in a head-to-head orientation with DEIN, a novel gene with stage specific expression in primary neuroblastoma (NB).  HAND2 and DEIN represent a gene pair that is closely linked by a bidirectional promoter in an evolutionary highly conserved manner. Expression of both genes in neuroblastoma is co-regulated by the asymmetrical activity of this promoter and modulated by the activity of two cis-regulatory elements acting as weak repressors (2).  Hand2 is developmentally regulated and is intranuclear in precursors but cytoplasmic in neurons. Hands is a 217 amino acid (26.5kDa) protein coded by Hand2 gene  located on chromosome 16p11.  Hand1 and 2 are expressed in adult heart and  are regulated by cardiac hypertrophy (3).

The Hand2 antibodies were generated using synthetic peptide derived from the from the N-terminal region of the Hand2 protein. The hand2-sepcific antibodies are affinity purified over immobilized antigen based affinity chromatography, and the purified immunoglobulins are stabilized in antibody stabilization buffer.  FabGennix Int. Inc., will provide limited quantities of antigenic blocking peptides and western blot positive control for Hand2 in ready-to-use buffer for Western blotting.  FabGennix Inc. will also conjugate antibodies with fluorescent probes upon request at a reasonable cost. FabGennix International Inc. also carries a number of antibodies to Transcription factors and RNA binding proteins, for a complete listing visit www.FabGennix.com. 

Immunogen:  Synthetic peptide taken from the C-terminal end of heart & neuronal-crest derivative protein 2.  The peptide was conserved in several other species.  The selected peptide was post-synthetically modified to achieve highest antigenicity before coupling to carrier protein using hetero bifunctional cross linker for immunogen preparation.          

Concentration:  Affinity purified Hand-201AP antibody (0.53-0.63mg/ml) was stabilized in antibody stabilization buffer for long-term storage.

Applications:  Hand-201AP antibody was tested in ELISA and western blotting applications at 1:500 dilution using PC-Hand2 samples.  Antibody dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions.  Application of this antibody in other protocols has not yet tested. WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC n.d. Not determined, does not necessarily exclude the use of this antibody in a protocol, investigators using this antibody in protocols other than listed above can request a complimentary sample of this antibody.

Reactivity: The antibody Hand-201AP labels a 25 kDa protein in PC-Hands 2 samples and in rodent heart. 

Protocols:  Standard protocol for various applications (WB, IMM, IHC) of this antibody can be obtained upon request. The specification sheet for Hand-201AP antibody will be supplied with each product.  FabGennix Inc., strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.  

Storage:   The antiserum is supplied in antibody stabilization buffer with 0.02% azide.  The purified antibodies are isolated from immobilized antigen affinity column.  The affinity pure antibodies are supplied as 0.5-1 mg/ml IgG in antibody stabilization buffer.   For long-term storage of keep at -20^oC.  FabGennix Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DiluOBuffer).  Working solutions of antibodies in DiluOBuffer should be filtered through 0.45m filter after every use for long-term storage.

 

 

For research use only, not for diagnostic or therapeutic use.