DYNC1LI1 anticorps (C-Term)

N° du produit ABIN2790248

Détail du produit anti-DYNC1LI1 anticorps

Antigène: DYNC1LI1 (Dynein, Cytoplasmic 1, Light Intermediate Chain 1 (DYNC1LI1))

Épitope: C-Term

Reactivité: Humain, Souris, Boeuf (Vache), Chien, Rat, Cheval, Cobaye, Porc, Lapin

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp DYNC1LI1 est non-conjugé

Application: Western Blotting (WB)

Séquence: AEDDQVFLMK LQSLLAKQPP TAAGRPVDAS PRVPGGSPRT PNRSVSSNVA

Homologie: Cow: 100%, Dog: 100%, Guinea Pig: 93%, Horse: 100%, Human: 100%, Mouse: 100%, Pig: 100%, Rabbit: 100%, Rat: 100%

Attributs du produit: This is a rabbit polyclonal antibody against DYNC1LI1. It was validated on Western Blot.

Purification: Affinity Purified

Immunogène: The immunogen is a synthetic peptide directed towards the C-terminal region of Human DYNC1LI1

Information d'application

Indications d'application: Optimal working dilutions should be determined experimentally by the investigator.

Commentaires:

Antigen size: 523 AA

Restrictions: For Research Use only

Independent Validation (WB)

Validation #100203 (Western Blotting)

by: VIB Medical Biotechnology Center, University of Gent

No.: #100203

Date: 16.02.2017

Antigène: DYNC1LI1

Numéro du lot: QC36279

Application validée: Western Blotting

Contrôle positif: A549 cells

Conclusion: ABIN2790248 recognizes DYNC1Li1 in A549 cell lysates.

Validation Images

DYNC1Li1 protein detection with ABIN2790248 after western blotting on nitrocellulose membrane. Cell lysates made from about 500.000 (1), 125.000 (2) or 25.000 (3) A549 cells were separated on a 10% acrylamide SDS-PAGE gel and blotted on a nitrocellulose membrane. DYNC1Li1 detection was performed using ABIN2790248 as primary antibody, followed by a secondary donkey-anti-rabbit-HRP antibody and ECL Western blotting substrate. M: marker

Protocole

Anticorps primaire: ABIN2790248

Anticorps secondaire: HRP linked rabbit IgG (from donkey) secondary antibody

Full Protocol:

• A549 Cells are grown in DMEM medium (Thermo Fisher scientific, 52100-047, lot 1779868X) supplemented with 10% serum (Gibco Life technologies, 10270, lot 41G3844K) and 2mM L-glutamine (Lonza, BE17-605F, lot 6MB149), at 37°C and 5% CO₂ in a 12ml volume on a 75cm² dish.
• Detach cells with trypsin/EDTA and centrifuge at 400xg for 5min.
• Wash cells gently with PBS and centrifuge at 400xg for 5min.
• Lyse cells in 200 µl cold lysis buffer (50mM Tris-HCl pH8.0, 150mM NaCl, 5mM EDTA, 1% NP40, protease inhibitor cocktail (Roche), 0.1% SDS) for 20min on ice.
• Denature total protein for 10min at 99°C in 30µl Laemmli-SDS sample buffer and subsequently separate them on a denaturing 10% SDS-PAGE gel.
• Transfer proteins onto nitrocellulose membrane (Perkin Elmer, NBA085G001EA, lot no A10064498) with a semi-dry Western blotting system (Hoefer, TE77X) for 1.5h at 1 mA/cm².
• Block the membrane with 4% non-fat milk powder in PBST (PBS with 0.1% Tween20) ON at 4°C.
• Incubation with primary DYNC1LI1 antibody (antibodies-online, ABIN2790248, lot QC36279) diluted 1:2000 in TBS-T buffer ON at 4°C.
• Wash membrane 4x with PBST.
• Incubation with HRP linked rabbit IgG (from donkey) secondary antibody (Amersham, NA934, lot no 9551642) diluted 1:5000 in TBS-T buffer for 1h at RT.
• Wash membrane 4x times with PBST.
• Reveal protein bands using ECL Western blotting substrate (Thermo fisher scientific, 32106, lot no RC227810) on a chemiluminescence imager (Amersham, 29083461).

Notes: The DYNC1LI1 antibody ABIN2790248 reveals a protein of the expected molecular weight of 56kDa in lysates of A549 cells. Multiple other protein bands at lower molecular weight were also detected with the ABIN2790248 antibody. DYNC1LI1 protein knockdown could unravel if these lower molecular weight bands originate from DYNC1Li1 protein degradation or from aspecific binding of the ABIN2790248 antibody to other human proteins.

Stockage

Format: Liquid

Concentration: Lot specific

Buffer: Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: -20 °C

Stockage commentaire: For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.

Détails sur DYNC1LI1

Antigène: DYNC1LI1 (Dynein, Cytoplasmic 1, Light Intermediate Chain 1 (DYNC1LI1))

Autre désignation: DYNC1LI1 (DYNC1LI1 Produits)

Synonymes: anticorps 1110053F02Rik, anticorps Dnclic1, anticorps LIC-1, anticorps Dncli1, anticorps DNCLI1, anticorps LIC1, anticorps DLCA, anticorps LIC5759, anticorps dynein cytoplasmic 1 light intermediate chain 1, anticorps Dync1li1, anticorps DYNC1LI1

Sujet: DYNC1LI1 acts as one of several non-catalytic accessory components of the cytoplasmic dynein 1 complex that are thought to be involved in linking dynein to cargos and to adapter proteins that regulate dynein function. Cytoplasmic dynein 1 acts as a motor for the intracellular retrograde motility of vesicles and organelles along microtubules. DYNC1LI1 may play a role in binding dynein to membranous organelles or chromosomes. DYNC1LI1 is probably involved in the microtubule-dependent transport of pericentrin. DYNC1LI1 is required for progress throuh the spindle assembly checkpoint. The phosphorylated form appears to be involved in the selective removal of MAD1L1 and MAD1L2 but not BUB1B from kinetochores.
Alias Symbols: DNCLI1, FLJ10219
Protein Interaction Partner: REL, BICD2, DYNC1I1, DCTN1, UBC, DYNLRB1, SERBP1, HNRNPA0, AKAP12, DYNLT3, RPS12, PSMD8, PSMC6, EIF3E, FLNA, DYNC1LI2, DYNC1I2, DYNC1H1, vpr, VCP, ZFR, TXNRD1, TKT, DYNLT1, RANGAP1, IKBKG, CUL3, Dynll1, tat, RAB4A, NUP85, PCNT,
Protein Size: 523

Poids moléculaire: 56 kDa

ID gène: 51143

NCBI Accession: NM_016141, NP_057225

UniProt: Q9Y6G9