Anti-Dynein, 74 kDa Intermedia

Anti-Dynein, 74 kDa Intermediate chains, cytoplasmic, clone 74.1

Eukaryotic cells rely on actin and microtubule-based protein "motors" to generate intracellular movements. These protein "motors" contain specialized domains that hydrolyse ATP to produce force and movement along a cytoskeletal polymer (actin in the case of myosin family and microtubules in the case of the kinesin family and dyneins). Dynein has been implicated in cytoplasmic motile funtions, including chromosomal movement, retrograde organelle and axonal transport, the endocytic pathway, and the organization of the Golgi apparatus. In all cell types, dynein has the same basic structures and is composed of two or three distinct heavy chains (~450 kDa), three intermediate chains (70-125 kDa), and at least four light chains (15-25 kDa).

• Axonemal dynein intermediate chain 1
• dynein intermediate chain DNAI1
• dynein, axonemal, intermediate chain 1
• dynein, axonemal, intermediate polypeptide 1
• immotile cilia syndrome 1

74.1

Reacts with all known forms of the Mr = 74,000 Dalton intermediate chain subunit (IC74) of mammaliam cytoplasmic dynein. The antigenic site has been localized to the first 60 amino acids of the amino terminal portion of the molecule (a region conserved in both IC74 genes).

• Immunocytochemistry
• Immunofluorescence
• Immunoprecipitation
• Western Blotting

Immunocytochemistry:
1:50-1:100 dilution from a previous lot was used. Reacts with cultured MDCK cells, NRR cells, N1E cells, and PTK-1 cells.

Immunoprecipitation:
10 μL of a previous lot per 0.5 gm tissue was used. The monoclonal cleanly immunoprecipitates the entire dynein complex stoichiometrically from TX-100 or NP-40 lysates (including the 530 kD heavy chain, the light intermediate chains and the light chains) from various tissues and cultured cell lines. If detergents such as SDS are used to prepare the lysates the only dynein subunits which are immunoprecipitated are the IC74 subunits (presumably because the dynein complex dissociates), and other contaminating proteins can be found in SDS immunoprecipitates.

Immunoblotting:
1:1,000-1:5000. Reacts with cultured MDCK cells, PC-12 cells, N1E cells, neurons, glia and other cultured cells.

Optimal working dilutions must be determined by end user.

• Human
• Mouse
• Rat
• Bovine
• Drosophila
• Sheep
• Fish
• Xenopus

Rat, mouse, bovine, sheep, fish, Drosophila and Xenopus. Does not react with squid. Not tested on chicken, nematodes or Dictystelium.

Squid

IgG2b

Bovine brain cytoplasmic dynein.

74 kDa Intermediate chains, cytoplasmic

74 kDa

HeLa cells, A431 cell lysate

Purified mouse monoclonal IgG2b in buffer containing 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative.

Stable for 6 months at 2-8°C in undiluted aliquots from date of receipt.

• DNAI1
• PCD
• ICS
• MGC26204
• CILD1

Routinely evaluated by Western Blot on A431 lysates.

Western Blot Analysis:
1:500 dilution of this lot detected DYNEIN on 10 μg of A431 lysates.

Cell Structure

Cytoskeleton

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Chemicon (Millipore)

1 mg/mL

Monoclonal Antibody

Purified

Mouse