DYNC1LI1 antibody (C-Term)

Catalog No. ABIN2790248

Product Details anti-DYNC1LI1 Antibody

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

Binding Specificity: C-Term

Reactivity: Human, Mouse, Cow, Dog, Rat, Horse, Guinea Pig, Pig, Rabbit

Host: Rabbit

Clonality: Polyclonal

Conjugate: This DYNC1LI1 antibody is un-conjugated

Application: Western Blotting (WB)

Sequence: AEDDQVFLMK LQSLLAKQPP TAAGRPVDAS PRVPGGSPRT PNRSVSSNVA

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

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

Purification: Affinity Purified

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

Application Details

Application Notes: Optimal working dilutions should be determined experimentally by the investigator.

Comment:

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: 02/16/2017

Antigen: DYNC1LI1

Lot Number: QC36279

Method validated: Western Blotting

Positive Control: A549 cells

Notes: 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

Full Methods

Primary Antibody: ABIN2790248

Secondary Antibody: 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).

Experimental 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.

Handling

Format: Liquid

Concentration: Lot specific

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

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freeze-thaw cycles.

Storage: -20 °C

Storage Comment: 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.

Target Details for DYNC1LI1

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

Alternative Name: DYNC1LI1 (DYNC1LI1 Products)

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

Background: 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

Molecular Weight: 56 kDa

Gene ID: 51143

NCBI Accession: NM_016141, NP_057225

UniProt: Q9Y6G9