itk (Cell Signaling Technology Inc)
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Structured Review
Cell Signaling Technology Inc
itk
Itk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/itk/ITK+Rabbit+mAb/bio_rxiv__64898__2026__01__28__701378-275-41-42
Average 94 stars, based on 6 article reviews
Itk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/itk/ITK+Rabbit+mAb/bio_rxiv__64898__2026__01__28__701378-275-41-42
Average 94 stars, based on 6 article reviews
itk - by Bioz Stars,
2026-10
94/100 stars
Images
1) Product Images from "C-terminus CD28 phosphorylation (Y218) modulates IL-2 secretion and antitumor effect of CAR-T cells"
Article Title: C-terminus CD28 phosphorylation (Y218) modulates IL-2 secretion and antitumor effect of CAR-T cells
Journal: bioRxiv
doi: 10.64898/2026.01.28.701378
Figure Legend Snippet: A . Top panel: Schematic representation of the protocol used to inhibit ITK activity. PSCA CAR-T cells were pre-treated for 2 hours with ITK inhibitors 1μM (BMS-509744, Ibrutinib or GNE-9822) or DMSO as control and then cocultured with HPAC WT cells for 0 or 10 minutes. Left panel: Representative Western blot of CD28 pY218 in CAR-T cells after stimulation with HPAC WT cells. Right panel: Normalized CD28 pY218 intensity with respect to CAR (CD3ζ). Significance was determined using one-way ANOVA. * = P<0.05. Each symbol represents an independent experiment from 3 different healthy donors. Data is represented as the mean ± standard deviation (SD). B. Top panel. Schematic representation of the experimental protocol. ITK expression was disrupted using CRISPR/Cas in Jurkat cells. WT and ITK-KO Jurkat cells were transduced to express a PSCA-specific CAR and cocultured with HPAC WT cells for 0 or 10 min. Left panel. Representative membrane of CD28 pY218 in CAR-T Jurkat cells after stimulation. Right panel. Normalized CD28 pY218 intensity quantified by densitometry (representative plot) and CD28 pY218 at 10 min post-stimulation. Significance was determined by paired t-test. ** = P<0.01. Each symbol represents one of 3 independent experiments, performed with different donor T cells.
Techniques Used: Activity Assay, Control, Western Blot, Standard Deviation, Expressing, CRISPR, Membrane
Figure Legend Snippet: A . Schematic representation of WT and PYRP mutant CAR constructs. In the PYRP mutant, alanine (A) 217 and serine (S) 220 were substituted with prolines (P) to generate a proline-rich domain. B . PSCA-targeted WT and PYRP CAR-T cells were cocultured with HPAC WT cells for 0, 1, 10, 30, or 60 min. CAR molecules were immunoprecipitated using protein-L coated beads and co-immunoprecipitated proteins were analyzed by Western blot. Upper panel: Representative Western blot results showing ITK present in CAR immunoprecipitate. CD3ζ staining of the CAR molecule was used as a bait control. Lower panel: Intensity of ITK with respect a total CAR (CD3ζ) was analyzed by densitometry (representative plot). C. PSCA WT and PYRP CAR-T cells were stimulated with HPAC WT cells for 10 min and phosphorylation of Y218 was analyzed by mass spectrometry. Bar charts represent the intensity of phosphorylated Y218 normalized to the total CAR signal. Significance was determined by one-way Anova. * = P<0.05. Data is represented as the mean ± standard deviation (SD). D and F. PSCA WT and PYRP CAR-T cells were cocultured with HPAC WT cells. Supernatant was collected after 24 hours and cytokine production was measured by ELISA. Significance was determined by one-way ANOVA. * = P<0.05, ** = P<0.01. Each symbol represents an independent experiment from 3 different healthy donors. Data is represented as the mean ± standard deviation (SD). E. PSCA WT and PYRP CAR-T cells were cocultured with HPAC WT cells. RNA was extracted after 24 hours and IL-2 mRNA production was evaluated using quantitative Time PCR. CAR mRNA was used as normalization control. Significance was determined by one-way Anova. ** = P<0.01, *** = P<0.001. Each symbol represents an independent experiment from 3 different healthy donors. Data is represented as the mean ± standard deviation (SD). G. % of change in tumor volume (compared to baseline) is represented (n=5). Representative example of two independent experiments. Significance was determined by permutation two sample t-test. * = P<0.05.
Techniques Used: Mutagenesis, Construct, Immunoprecipitation, Western Blot, Staining, Control, Phospho-proteomics, Mass Spectrometry, Standard Deviation, Enzyme-linked Immunosorbent Assay
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