kindlin-2 antibody | knockout validation | Millipore MAB2617

This is a knockout-validated antibody summary, based on the publications "Kindlin-2 cooperates with talin to activate integrins and induces cell spreading by directly binding paxillin", as cited below [1], "Genetic Compensation Triggered by Mutant mRNA Degradation" for western blot with transfected Fermt2 (figure e2e) [2], and "Kindlin-2 Suppresses Transcription Factor GATA4 Through Interaction With SUV39H1 to Attenuate Hypertrophy" for immunohistochemistry (figure 5e) [3]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

kindlin-2 antibody | knockout validation | Millipore MAB2617 figure 1
Figure 1. Western blot of kindlin-2 in control, talin-KO (TlnKo), kindlin-KO (KindKo) and keratinocyte (Kerat.) cells. GAPDH served as loading control.
Antibody information

Mouse monoclonal IgG1κ

Company: Millipore, now part of MilliporeSigma

Antibody: kindlin-2

Catalog number: MAB2617

Summary: Mouse monoclonal IgG1κ against GST-tagged recombinant protein corresponding to human Kindlin-2. Reacts with human, and is predicted to react with bovine (based on 100% sequence homology), and mouse and rat (98% sequence homology). Suitable for western blot and immunocytochemistry.

Validation Method

Western blot | Immunocytochemistry

Sample

WB: kindlin-1 and -2 double knockout fibroblasts derived from mouse kidneys.

IC: kindlin-1 and -2 double knockout fibroblasts derived from mouse kidneys, fixed in –20°C cold acetone-methanol and permeabilized with –20°C cold methanol.

Blocking agent

IC: 1 hr at RT in PBS supplemented with 0.1% Triton X-100 (v/v) and 3% BSA (w/v)

Primary incubation

WB: 1:1000

IF: 1:500

Secondary incubation

WB: goat anti-mouse HRP (172-1011) (BioRad), 1:10,000

IC: 1:500 goat anti-mouse Alexa 488 (A11029, Invitrogen) in blocking buffer in the dark

Detection

IC: LSM 780 confocal microscope (Zeiss, Germany) equiped with a 100×/NA 1.46 oil objective and with a DMIRE2-SP5 confocal microscope (Leica, Germany) equiped with a 40×/NA 1.25 or 63×/NA 1.4 oil objective using Leica Confocal software (version 2.5 build 1227).

References
  1. Theodosiou M, Widmaier M, Böttcher R, Rognoni E, Veelders M, Bharadwaj M, et al. Kindlin-2 cooperates with talin to activate integrins and induces cell spreading by directly binding paxillin. elife. 2016;5:e10130 pubmed publisher
  2. El Brolosy M, Kontarakis Z, Rossi A, Kuenne C, Günther S, Fukuda N, et al. Genetic compensation triggered by mutant mRNA degradation. Nature. 2019;: pubmed publisher
  3. Qi L, Chi X, Zhang X, Feng X, Chu W, Zhang S, et al. Kindlin-2 suppresses transcription factor GATA4 through interaction with SUV39H1 to attenuate hypertrophy. Cell Death Dis. 2019;10:890 pubmed publisher