Cf inp d281a

Manufactured by Addgene

The CF-Inp(D281A) is a lab equipment product. It is a variant of the Inp protein, with the aspartic acid at position 281 replaced with alanine. The core function of this product is to serve as a research tool for studying protein-protein interactions and enzyme activity.

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Lab products found in correlation

Gfp plcδ ph, by Addgene (1 mentions) Ph btk gfp, by Addgene (1 mentions) Akt ph gfp, by Addgene (1 mentions)

2 protocols using cf inp d281a

Constructing LDR-T2A-Inp54p Fusion Protein

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To generate LDR-T2A-Inp54p, Lyn₁₁-targeted FRB (LDR) (plasmid #20147) and CF-Inp54p (plasmid #20155) were obtained from Addgene. As a backbone vector, pCDH-EF1-MCS-T2A-copGFP vector was used. The PCR fragment containing the LDR plus T2A sequence was digested with XbaI and PstI and ligated into the backbone vector between XbaI and PstI. Subsequently, the PCR fragment of CF-Inp54p was digested with PstI and SalI and was inserted into the LDR-T2A sequence containing vector. eCFP was replaced by eGFP amplified from EGFP-N1. In the LDR-T2A-Inp54p construct, Inp54p was superseded by the PCR fragment of Inp54p(D281A) from CF-Inp(D281A) (Addgene, plasmid 20156). PH-GFP (plasmid #21179) and PH-mCherry (plasmid #36075) were purchased from Addgene.

Kim S.J., Jeong M.J., Jo H.J., Jung J.H., Kaang B.K., Choi Y.B, & Kim J.H. (2017). Identification of postsynaptic phosphatidylinositol-4,5-bisphosphate (PIP2) roles for synaptic plasticity using chemically induced dimerization. Scientific Reports, 7, 3351.

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Protein Labeling and Imaging Techniques

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Mutations were introduced into DOC2B-GFP N2 ^{7 (link)}, C2AB-GFP N2 or C2B-GFP N2 ^{6 (link)}, using site-directed mutagenesis PCR. Botulinum toxins BoNT/C and BoNT/E were a generous gift from the laboratory of Prof. Ilana Lotan (Tel-Aviv University). mRFP-syx1A and SNAP25-YFP were a kind gift from Prof. Edward Stuenkel (University of Michigan). R-GECO was a generous gift from the laboratory of Prof. Robert Campbell ^{31 (link)}. Lact-C2-GFP was a gift from Prof. Sergio Grinstein (addgene #22852). For PI manipulations using the rapamycin dimerization system, the PH domain of PLCδ1, PLCδ-PH-GFP (addgene #21179), CF-Inp (addgene #20155), CF-Inp^D281A (addgene #20156) and Lyn11-FRB (addgene #20147) were a gift from Prof. Tobias Meyer. CF-PIPK, CF-PIPKi, mcherry-iSH were a generous gift from the laboratory of Prof. Takanari Inoue (Johns Hopkins University). PLCδ-PH-mKate was a kind gift from Prof. Thomas Martin (University of Wisconsin-Madison). Rab7-FRB (addgene #51613), PH-Akt-GFP (addgene #51465), and PH-Btk-GFP (addgene #51463) were a gift from Prof. Tamas Balla. NPY-mRFP was a kind gift from Prof. Matthijs Verhage (Vrije University).

Michaeli L., Gottfried I., Bykhovskaia M, & Ashery U. (2017). Phosphatidylinositol (4,5)-bisphosphate targets DOC2B to the plasma membrane. Traffic (Copenhagen, Denmark), 18(12), 825-839.

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