Anti-Kindlin 2 antibody

• IP

Unknown

Immunoprecipitation - Anti-Kindlin 2 antibody (AB74030)

Kindlin 2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Kindlin 2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab74030.
Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band : 78kDa; Kindlin 2

All lanes:

Immunoprecipitation - Anti-Kindlin 2 antibody (ab74030)

Predicted band size: 78 kDa

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• WB

Lab

Western blot - Anti-Kindlin 2 antibody (AB74030)

Lanes 1-3 : Merged signal (red and green). Green - ab74030 observed at 78 kDa. Red - loading control ab8245 observed at 36 kDa.

ab74030 Anti-Kindlin 2 antibody was shown to specifically react with Kindlin 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265247 (knockout cell lysate ab257436) was used. Wild-type and Kindlin 2 knockout samples were subjected to SDS-PAGE. ab74030 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-Kindlin 2 antibody (ab74030) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FERMT2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FERMT2 (Kindlin 2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-fermt2-kindlin-2-knockout-hela-cell-line-ab265247'>ab265247</a>)

Lane 3:

Saos-2 cell 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: 78 kDa

Observed band size: 78 kDa

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• WB

AbReview19066****

Western blot - Anti-Kindlin 2 antibody (AB74030)

The kindlin 2 antibody (ab74030) was tested in lysate obtained from CHO cells either expressing endogenous levels of kindlin 2 (lane1)and transfected CHO cells in which flag-tagged kindlin2 was overexpressed (lane 2). A specific band was at the expected molecular weight of approximately 72 kDa (+/- Flag-tag). ß-actin was used to show loading in the two lanes.

All lanes:

Western blot - Anti-Kindlin 2 antibody (ab74030)

Predicted band size: 78 kDa

false

This image is courtesy of an abreview submitted by Ana Kasirer-Friede,UCSD , United States

• WB

Project

Western blot - Anti-Kindlin 2 antibody (AB74030)

All lanes:

Western blot - Anti-Kindlin 2 antibody (ab74030) at 1 µg/mL

Lane 1:

HT 1080 (Human fibrosarcoma) Whole Cell Lysate at 10 µg

Lane 2:

SAOS-2 (sarcoma osteogenic) Whole Cell lysate at 10 µg

Lane 3:

Human testis tissue lysate - total protein (<a href='/en-us/products/unavailable/human-testis-tissue-lysate-total-protein-ab30257'>ab30257</a>) at 10 µg

Lane 4:

HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 78 kDa

Observed band size: 78 kDa

false

Exposure time: 8min