Cells were seeded in 12 mm cover slips. After the indicated treatments, cells were fixed for 20 min at RT with 4% paraformaldehyde and permeabilized 0.5% NP-40 in PBS containing 0.5% BSA (Bovine serum albumin) for 10 min. After blocking for 1 h with 10% FBS in PBS containing 0.5% BSA, cells were incubated with the indicated antibodies: Anti-HA (Cat: 901514, Biolegend or Cat: 9110, Abcam) and anti-Abl 1:1000 (Sigma, Cat. A5844) overnight at 4 °C following by Duolink manufacturer´s instructions (Duolink®, Sigma-Aldrich). Images were acquired by confocal microscopy (Nikon C2 plus) using a 60X oil objective lens stacking the images every 0.5 µm to cover all the image of interest. Stack images were deconvoluted using Huygens and ImageJ. Stack deconvolved images were reduced to one dimension by sumslices function (ImageJ). Colocalization was performed in thresholded and masked images were used to determine Manders/Pearson’s index was calculated with ImageJ plugin.
Anti abl
Manufactured by Merck Group
Anti-Abl is a laboratory equipment product designed to detect and analyze Abl kinase activity. It is a research tool used in the study of cellular signaling pathways and protein interactions.
Automatically generated - may contain errors
Lab products found in correlation
Type 1 collagen, by BD (2 mentions)
Protein g agarose bead, by Santa Cruz Biotechnology (2 mentions)
X tremegene hp reagent, by Roche (2 mentions)
Anti actin, by Merck Group (2 mentions)
Anti py221 crkii, by Merck Group (2 mentions)
Anti crkii, by Merck Group (2 mentions)
Human egf, by Thermo Fisher Scientific (2 mentions)
Anti flag, by Merck Group (2 mentions)
C2 plus, by Nikon (1 mentions)
Anti ha, by BioLegend (1 mentions)
3 protocols using anti abl
1
Colocalization Analysis of HA and Abl Proteins
2
Crk Signaling Pathway Dissection
3
Crk Signaling Pathway Dissection
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The following antibodies were used: anti-pY221 CrkII (Sigma SAB4503813), anti-CrkII (Sigma SAB1405658), anti-actin (Sigma A2066), anti-Flag (Sigma F1804) and anti-Abl (Sigma SAB4501043). Protein G agarose beads were purchased from Santa Cruz Biotechnology, HRP-conjugated goat anti-mouse and goat anti-rabbit were purchased from Jackson Immunologicals and Type I Collagen was purchased from BD. Anti-Crk (RF-51) anti-sera was prepared as described previously51 (link). Transfections were performed with the Xtremegene HP reagent according to the manufacturers’ protocol (Roche). Human EGF was purchased from Life Technologies.
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