- PA5-25750 - Invitrogen Antibodies RASGRP1 antibody

Submitted references Loss of RASGRP1 in humans impairs T-cell expansion leading to Epstein-Barr virus susceptibility.

Winter S, Martin E, Boutboul D, Lenoir C, Boudjemaa S, Petit A, Picard C, Fischer A, Leverger G, Latour S
EMBO molecular medicine 2018 Feb;10(2):188-199

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis using a RASGRP1 polyclonal antibody (Product # PA5-25750) in HepG2 cell lysates (35 µg per lane).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis in formalin-fixed, paraffin-embedded human brain tissue using a RASGRP1 polyclonal antibody (Product # PA5-25750), followed by HRP-conjugated secondary antibody and DAB staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry analysis of HepG2 cells using a RASGRP1 polyclonal antibody (Product # PA5-25750) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Identification of a homozygous loss-of-function mutation in RASGRP1 in two siblings with Hodgkin lymphoma and defective immunity to EBV Pedigree of the family in which the c.1910_1911insAG mutation in RASGRP1 was identified. The arrow indicates the proband (P1.1) who was analyzed by WES. EBV load in the blood of patient P1.1 is shown as the number of EBV copies detected by PCR at different time points (black circles). Arrows correspond to the anti-CD20/rituximab treatments received by the patient with the age (year, y; month, m) of patient at the time of the treatment. Schematic representation of intron-exon organization of the RASGRP1 gene and its correspondence at protein level with the different domains of RASGRP1 shown: the Ras exchanger motif (REM), the Ras-guanine exchange factor (RasGEF), the EF-hand, the C1, and the bZIP domains. The mutation is indicated by an arrow at gene and protein levels. DNA electropherograms of the family showing the g.38786931_38786932insAG mutation in P1.1 and P1.2 that is shown in the box. Expression of RASGRP1 transcript in T-cell blasts of healthy control and the patient P1.1 (Pat.). The relative expression of full-length RASGRP1 transcript was examined by qRT-PCR in T-cell blasts of a healthy control and P1.1. Fourfold serial dilutions of cDNAs (1, 0.5, 0.25, and 0.12) were used for amplification of each transcript after quantitation. Base pair markers are shown on the left. PCR products were verified by sequencing showing th

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 RASGRP1 is required for T-cell expansion in response to CD3-TCR activation Rescue of T-cell proliferation by expression of wild-type RASGRP1 in RASGRP1-deficient T cells. Immunoblots for RASGRP1 and actin as a loading control of control (Ctr.) and RASGRP1-deficient T cells of patient P1.1 (Pat.) transduced by empty or wild-type RASGRP1-containing vector with mCherry gene reporter at day 14 of culture. Proliferation of control (Ctr.) and RASGRP1-deficient T cells of patient P1.1 (Pat.) transduced by empty or wild-type RASGRP1-containing vector. Representative histograms of CellTrace violet dye dilution showing the cell divisions of mCherry-positive (mCherry + ) and mCherry-negative (mCherry - ) cells after stimulation with two concentrations of anti-CD3 antibody, with histograms in gray corresponding to unstimulated cells (unstim.). Curves of the percentage of mCherry-positive transduced cells after repeated stimulations indicated by gray arrows with anti-CD3 (upper panels) or anti-CD3/CD28 (lower panels) antibodies at different days in long-term expansions. Data information: Representative data from two independent experiments. Source data are available online for this figure.

PA5-25750

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