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

Cytoplasmic dynein 1 intermediate chain 1 (UniProt Q29RQ3; also known as Cytoplasmic dynein intermediate chain 1, Dynein intermediate chain 1, cytosolic, DH IC-1) and Cytoplasmic dynein 1 intermediate chain 2 (UniProt Q0III3; also known as Cytoplasmic dynein intermediate chain 2, Dynein intermediate chain 2, cytosolic, DH IC-2) are encoded by the DYNC1I1 gene (Gene ID 613724) and DYNC1I2 gene (Gene ID 526329) in bovine, respectively. Cytoplasmic dynein complexes are microtubules-associated retrograde transport motors. Cytoplasmic dynein 1 is the more abundant motor in cells, while cytoplasmic dynein 2 takes part in intraflagellar transport. Cytoplasmic dynein 1 is a multisubunit complex of ∼1.5 MDa composed of a homodimer of heavy chains (encoded by DYNC1H1), two intermediate chains (encoded by DYNC1I1 and DYNC1I2), light-intermediate chains (encoded by DYNC1LI1, DYNC1LI2) and light chains (encoded by DYNLT1, DYNLT3, DYNLRB1, DYNLRB2, DYNLL1, DYNLLl2). The heavy chain homodimer acts as the dynein complex core and binds microtubules to enable ATP-dependent cytoplasmic dynein movement. The other dynein subunits associate as homodimers and play regulatory roles in maintaining the complex stability, modulating its activity, and mediating its interaction with accessory and cargo proteins. The intermediate chain proteins (IC1 & IC2) are involved in cargo binding and specificity, either alone or via interaction with the dynactin complex. IC1 and IC2 share 69% protein identity. They interact with the dynein light chains and the p150 subunit of dynactin at the N-terminus and with the heavy chains through WD40 repeats at the intermediate chain C-terminus.

Not tested on chicken, nematodes or Dictystelium.

Anti-Dynein Antibody, 74 kDa Intermediate chains, cytoplasmic, clone 74.1 detects level of Dynein & has been published & validated for use in IC, IF, IP & WB.

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.

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.

~74 kDa observed.

Format: Purified

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.

Control
HeLa cells, A431 cell lysate

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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