Anti-DYNLL1/PIN antibody [EP1660Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Immunohistochemical staining of paraffin embedded human liver using unpurified ab51603 (1/100).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling DYNLL1/PIN (red) with purified ab51603 at a 1/2300 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• ICC/IF

AbReview16748****

Immunocytochemistry/ Immunofluorescence - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Unpurified ab51603 staining DLC8 in mouse kidney cells cells by ICC/IF (immunocytochemistry/immunofluorescence. Cells were fixed with methanol, permeabilized with 0.1% Triton and blocked with 1% milk for 1 hour at room temperature. The sample was incubated with primary antibody (1/400; 1% milk in PBS) for 16 hours at 4°C. An Alexa Fluor^®488-conjugated Goat polyclonal to rabbit IgG (1/1000) was used as secondary antibody.

This image is courtesy of an anonymous Abreview.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling DYNLL1/PIN with Purified ab51603 at 1 : 100 dilution (6.7μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1 : 1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling DYNLL1/PIN with Purified ab51603 at 1 : 500 dilution (1.34 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IP

PubMed

Immunoprecipitation - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Unpurified ab51603 used in IP.SKAP and Astrin form a complex. (A, left) Silver-stained gels showing a one-step IP of GFPLAP-Astrin, GFPLAP-SKAP, or GFPLAP-LC8. (A, right) Data from the mass spectrometric analysis of the purifications indicating the percent sequence coverage from each IP. (B) Silver-stained gel showing the purification of FLAG-SKAP from chicken DT40 cells relative to controls. The indicated proteins were identified by excising them from a gel and analyzing them by mass spectrometry.

All lanes:

Immunoprecipitation - Anti-DYNLL1/PIN antibody [EP1660Y] (ab51603)

Predicted band size: 10 kDa

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Image from Schmidt JC et al. J Cell Biol. 2010 Oct 18;191(2):269-80. Epub 2010 Oct 11 Fig 2. DOI 10.1083/jcb.201006129.

• IP

Unknown

Immunoprecipitation - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

ab51603 (purified) at 1 : 30 dilution (2ug) immunoprecipitating DYNLL1/PIN in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.

Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab51603 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab51603 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-DYNLL1/PIN antibody [EP1660Y] (ab51603)

Predicted band size: 10 kDa

false

• WB

Unknown

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Lanes 1-3 : Merged signal (red and green). Green - ab51603 observed at 10 kDa. Red - loading control ab8245 observed at 36 kDa.

ab51603 Anti-DYNLL1/PIN antibody [EP1660Y] was shown to specifically react with DYNLL1/PIN in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265265 (knockout cell lysate ab257414) was used. Wild-type and DYNLL1/PIN knockout samples were subjected to SDS-PAGE. ab51603 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (ab51603) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

DYNLL1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human DYNLL1 (PIN) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-dynll1-pin-knockout-hela-cell-line-ab265265'>ab265265</a>)

Lane 3:

Mouse testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 10 kDa

Observed band size: 10 kDa

false

• WB

Lab

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (ab51603) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

Mouse testis lysates at 20 µg

Lane 3:

Rat testis lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 10 kDa

Observed band size: 10 kDa

false

• WB

AbReview54133****

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

Lymphnodes were dissociated in PBS 2% FBS. Cell suspensions filtered through 70 μm and 40 μm cell strainers, and 300 x g pellets were lysed in modified RIPA buffer (150 mM NaCl, 20 mM Tris pH7.4, 1 mM EDTA, 1 mM EGTA, 10 mM NaF, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 1 mM PMSF, 1 x protein inhibitor cocktail (Sigma)).

All lanes:

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (ab51603) at 1/5000 dilution

Lane 1:

Primary mouse Mb1-Cre control Eµ-Myc B cell lymphoma (lysate of whole lymphnode)

Lane 2:

Primary mouse Mb1-Cre DYNLL1/PIN-conditional knockout Eµ-Myc B cell lymphoma (lysate of whole lymphnode)

Secondary

All lanes:

HRP conjugated polyclonal goat IgG at 1/5000 dilution

Predicted band size: 10 kDa

Observed band size: 10 kDa

false

Exposure time: 10min

This image is courtesy of an abreview submitted by Dr. Jörg Heierhorst

• WB

Unknown

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (AB51603)

All lanes:

Western blot - Anti-DYNLL1/PIN antibody [EP1660Y] (ab51603) at 1/10000 dilution

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 10 kDa

Observed band size: 10 kDa

false