Gst b 14

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The GST (B-14) is a recombinant protein purification product offered by Santa Cruz Biotechnology. It is a form of the Glutathione S-Transferase (GST) protein, which is commonly used in affinity chromatography for the purification of recombinant fusion proteins. The core function of the GST (B-14) is to serve as an affinity tag that can be attached to target proteins, allowing for their selective capture and isolation from complex mixtures.

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Lab products found in correlation

P53 do 1, by Santa Cruz Biotechnology (3 mentions) Gapdh 6c5, by Santa Cruz Biotechnology (3 mentions) Cip p4978, by Merck Group (2 mentions) Myc 9e10, by Santa Cruz Biotechnology (2 mentions) Ha 12ca5, by Roche (2 mentions) Hif1α h1α67, by Santa Cruz Biotechnology (2 mentions) Nedd8 a 812, by R&D Systems (2 mentions) β actin, by Merck Group (2 mentions) Secondary peroxidase labeled anti mouse or anti rabbit immunoglobulin g antibodies, by Jackson ImmunoResearch (1 mentions) Lc3b d11, by Cell Signaling Technology (1 mentions) Anti flag m2 antibody, by Agilent Technologies (1 mentions) Image labtm software, by Bio-Rad (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Il 1β af 401 na, by R&D Systems (1 mentions) Rm4 5, by Thermo Fisher Scientific (1 mentions) β actin ac 15, by Merck Group (1 mentions) Ra3 6b2, by Thermo Fisher Scientific (1 mentions) Ha f 7, by Santa Cruz Biotechnology (1 mentions) Ha c29f4, by Cell Signaling Technology (1 mentions) Streptavidin hrp, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Anti-GST (B-14, (9 citations) Mouse anti-GST (B-14) (5 citations) Anti-GST clone B-14 (3 citations) Anti‐GST mAb B‐14 (2 citations) GST-tag (B-14) mouse monoclonal (sc-138) (2 citations)

13 protocols using gst b 14

Quantitative Western Blot Analysis of Cellular Signaling

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Cells were harvested, fractionated and transferred onto nitrocellulose membranes as described previously (Kim et al., 2015 (link)). Antibodies to STING (D2P2F), phospho-TBK1 (D52C2) and LC3B (D11) were purchased from Cell Signaling Technology. An anti-FLAG M2 antibody (#200474) was purchased from Agilent technologies. Antibodies to tubulin (B-5-1-2) and TBK1 (AOW9) were purchased from Sigma–Aldrich and EMD Millipore, respectively. Anti-HCMV IE (CH160) antibody was purchased from Virusys. Antibodies to IE86 (12E2), c-Myc (9E10) and GST (B-14) were purchased from Santa Cruz Biotechnology. Secondary peroxidase-labeled anti-mouse or anti-rabbit immunoglobulin G antibodies were purchased from Jackson ImmunoResearch. The signal intensity of protein bands were quantitated using Image Lab^TM software (Bio-Rad Laboratories).

Kim J.E., Kim Y.E., Stinski M.F., Ahn J.H, & Song Y.J. (2017). Human Cytomegalovirus IE2 86 kDa Protein Induces STING Degradation and Inhibits cGAMP-Mediated IFN-β Induction. Frontiers in Microbiology, 8, 1854.

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Molecular Mechanisms of PKC Signaling Regulation

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CIP (P4978) and DMSO were purchased from Sigma-Aldrich. PKCα (C2-4) inhibitor peptide (17478) was purchased from Cayman Chemical. Flag-PKCα, -β, -δ, and -ξ plasmids were a generous gift from Dr. D. Zhou (Xiamen University, China). The GST-tagged GRA7 and truncated mutant genes were described previously [17 (link)]. V5-tagged AC or AU1-PLD1 and truncated mutant genes were cloned into the XbaI and BamHI sites in pcDNA3.0. All constructs were sequenced using an ABI PRISM 377 automatic DNA sequencer to verify 100% correspondence with the original sequence. Specific antibodies against phospho-(Thr147)-PLD1 (3831), phospho-(Ser561)-PLD2 (3834), PLD1 (3832), PLD2 (13904), PKCα (2056), PKCγ (43806), and NLRP4 (12421) were purchased from Cell Signaling Technology. Antibodies specific for actin (I-19), ASC (N-15-R), IL-18 (H-173-Y), TRAF6 (H-274), caspase-1 p10 (M-20), Rab5 (D-11), Rab7 (H-50), LAMP1 (E-5), LAMP2 (H4B4), Tubulin (B-5-1-2), Calnexin (H-70), FACL4 (N-18), VDAC (B-6), His (His17), V5 (C-9), Flag (D-8), and GST (B-14) were purchased from Santa Cruz Biotechnology. AU1 (GTX23402) and PKCβI (A10-F) were purchased from GenenTex and Antibodies-online Inc., respectively. IL-1β (AF-401-NA) and NLRP3 (AG-20B-0014) were from R&D Systems and Adipogen, respectively.

Koh H.J., Kim Y.R., Kim J.S., Yun J.S., Jang K, & Yang C.S. (2017). Toxoplasma gondii GRA7-Targeted ASC and PLD1 Promote Antibacterial Host Defense via PKCα. PLoS Pathogens, 13(1), e1006126.

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Western Blot and Flow Cytometry Analysis

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The following Abs were used for Western blot analyses or flow cytometry analysis: p53 (DO-1, Santa Cruz), p21 (DSC60, Cell Signaling Technology, Danvers, MA), β-actin (AC-15, Sigma-Aldrich, St. Louis), His (Invitrogen), GST (B-14, Santa Cruz), HA (F-7, Santa Cruz), HA (C29F4, Cell Signaling Technology), Myc (9E10, Santa Cruz), Ab against mouse DD1α (MH5A, BioLegend), Ab against human PD-1 (J116, eBioscience), Ab against human PD-L1 (M1H1, eBioscience), CTLA4 (C-19, Santa Cruz), ICOSL (LifeSpan BioSciences), Ab against human CD14 APC (61D3, eBioscience), Ab against mouse F4/80 APC (BM8, eBioscience), Ab against mouse CD45R/B220 (RA3–6B2, eBioscience), Ab against human CD4-APC (OKT4, eBioscience), Ab against mouse CD4-APC (RM4–5, eBioscience), Ab against human CD8-APC (SK1, eBioscience) and Ab against mouse CD8a⁺-APC (53–6.7, eBioscience). Rabbit polyclonal and mouse monoclonal Ab against DD1α was raised against human DD1α (amino acids 33–311) as an immunogen. Beads immobilized with Abs against HA (Roche) or Myc (9B11, Cell Signaling) were used for immunoprecipitations. Recombinant human PD-1–Ig, mouse PD-1–Ig, human PD-L1–Ig, mouse PD-L1–Ig, and control Ig proteins were from R&D Systems.

Yoon K.W., Byun S., Kwon E., Hwang S.Y., Chu K., Hiraki M., Jo S.H., Weins A., Hakroush S., Cebulla A., Sykes D.B., Greka A., Mundel P., Fisher D.E., Mandinova A, & Lee S.W. (2015). Control of signaling-mediated clearance of apoptotic cells by the tumor suppressor p53. Science (New York, N.Y.), 349(6247), 1261669.

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Protein Extraction and Phosphorylation Analysis

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Yeast cell pellets (∼1.5–6.0 OD units) were suspended in 200 µl of 20% trichloroacetic acid, kept on ice for 15 min, and centrifuged at 15,000 g for 5 min. The pellets were washed with 1 ml of cold acetone, dried at room temperature, and suspended in (OD units × 50) µl of SDS sample buffer by mixing at 65°C for 10 min followed by cell disruption at room temperature using FastPrep-24 (MP Biomedicals) and 0.5-mm YZB zirconia beads (Yasui Kikai). These samples were boiled for 3 min and centrifuged at 15,000 g for 1 min; the supernatants were used for immunoblotting analysis (Nakatogawa and Ohsumi, 2012 (link)). To separate phosphorylated and unphosphorylated forms of nontagged Atg19, the Anderson gel system (10% acrylamide and bis-acrylamide at 77:1) was used (Anderson et al., 1973 (link)). Monoclonal antibodies against GFP (11814460001; Roche), AID (BioROIS), GST (B-14; Santa Cruz Biotechnology, Inc.), HA (3F10; Roche), and Myc (9E10; Santa Cruz Biotechnology, Inc.) sequences were used for detection of tagged proteins. The monoclonal antibody against Pgk1 was purchased from Invitrogen. The phosphate-binding reagent Biotinylated Phos-tag (Wako Pure Chemical Industries) was used to detect protein phosphorylation in combination with streptavidin-HRP (Invitrogen).

Tanaka C., Tan L.J., Mochida K., Kirisako H., Koizumi M., Asai E., Sakoh-Nakatogawa M., Ohsumi Y, & Nakatogawa H. (2014). Hrr25 triggers selective autophagy–related pathways by phosphorylating receptor proteins. The Journal of Cell Biology, 207(1), 91-105.

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Protein Immunoblotting Analysis

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Whole cell lysates, immunoprecipitated proteins, pulldown assays, and in vitro reactions were all subjected to SDS-PAGE and transferred to PVDF membrane. Western blots for HA (12CA5, Roche), p53 (DO-1, Santa Cruz), Mdm2 (2A10, 2A9, 4B2), VHL (VHL40, FL-181, Santa Cruz), GST (B-14, Santa Cruz), HIF1α (H1α67, Santa Cruz), Ac-p53 K373/382 (Upstate Signaling), GAPDH (6C5, Santa Cruz), and NEDD8 (A-812, Boston Biochem) were performed according to the manufacturers’ protocols.

Wolf E.R., Mabry A.R., Damania B, & Mayo L.D. (2020). Mdm2 mediated neddylation of pVHL blocks the induction of anti-angiogenic factors.. Oncogene, 39(29), 5228-5239.

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Protein Immunoblotting Analysis

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Wolf E.R., Mabry A.R., Damania B, & Mayo L.D. (2020). Mdm2 mediated neddylation of pVHL blocks the induction of anti-angiogenic factors.. Oncogene, 39(29), 5228-5239.

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Investigating CTR-p18 Protein Interaction

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Glutathione S-transferase (GST) pulldown analysis was performed as described previously (22 (link)). Briefly, pGST [pEG(KT)], pGST-CTR (pBDG1496) and pAUG1p18 (pBDG1608; pAUG1p18/TRP1) or psAUG1 [(deleted C-DR p18) pBDG1612; psAUG1/TRP1] were introduced into DG3582 by transformation. Cells were grown in SC-Ura-Trp + 2% Galactose for 2 days at 22°C after pre-culturing in SC-Ura-Trp + 2% Raffinose overnight at 30°C. To determine if the CTR interacts with p18, GST-complexes were analyzed by immunoblotting using mouse monoclonal antibody GST/B-14 (Santa Cruz Biotech) or rabbit polyclonal p18 antibody (22 (link)).

Nishida Y., Pachulska-Wieczorek K., Błaszczyk L., Saha A., Gumna J., Garfinkel D.J, & Purzycka K.J. (2015). Ty1 retrovirus-like element Gag contains overlapping restriction factor and nucleic acid chaperone functions. Nucleic Acids Research, 43(15), 7414-7431.

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Antibody Production and Characterization

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In the present study, we produced monoclonal anti-βPix-SH3 and polyclonal anti-βPix-GBD antibodies against purified GST-SH3 and GST-GBD proteins. In addition, we procured antibodies against Dyn2 (C-18, #sc-6400; Santa Cruz Biotechnology, Dallas, TX, USA), E-cadherin (24E10, #3195; Cell Signaling Technology, Danvers, MA, USA), MT1-MMP (L-15, #sc-12367; Santa Cruz Biotechnology), GAPDH (6C5, #sc-32233; Santa Cruz Biotechnology), Cortactin (H-191; #sc-11408; Santa Cruz Biotechnology), GST (B-14, #sc-138; Santa Cruz Biotechnology), c-Myc (9E10, #sc-40; Santa Cruz Biotechnology), FLAG (M2, #F1804; Sigma-Aldrich, St. Louis, MO, USA), GFP (B-2, #sc-9996; Santa Cruz Biotechnology), Rac1 (#610651; BD Transduction Laboratory, San Jose, CA, USA), and phosphotyrosine (4G10; #05-321; Millipore, Burlington, MA, USA). In addition, epidermal growth factor (EGF, E9644; Sigma-Aldrich), PP2, a Src kinase inhibitor (#529573; Calbiochem, La Jolla, CA, USA), poly L-lysine hydrobromide (#P6282; Sigma-Aldrich), fibronectin (#F2006; Sigma-Aldrich) and Matrigel (#354234; Corning, Corning, NY, USA) were used.

Keum S., Yang S.J., Park E., Kang T., Choi J.H., Jeong J., Hwang Y.E., Kim J.W., Park D, & Rhee S. (2021). Beta-Pix-dynamin 2 complex promotes colorectal cancer progression by facilitating membrane dynamics. Cellular Oncology (Dordrecht), 44(6), 1287-1305.

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Antibody Characterization for Stem Cell Markers

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The antibodies used were: policlonal VRK1 (VC) and monoclonal VRK1 (1B5 or 1F6)64 (link). VRK1 (HPA000660, Sigma). Sox2 (D6D9, Cell Signaling; Berverly, MA,; E4, Santa Cruz; Santa Cruz, CA; Y-17, Santa Cruz,). Oct4 (2840, Cell Signaling), Nanog (D73G4, Cell Signaling), N-Cadherin (H-63, Santa Cruz), Vimentin (RV202, Abcam; Cambridge, UK). Flag epitope (Sigma-Aldrich, monoclonal M5; Sigma-Aldrich, polyclonal F7425). HA epitope (Santa Cruz, F-7; Sigma-Aldrich, H6908), myc epitope (06–549, Millipore; Billerica, MA; 05–724, Millipore), GST (B-14, Santa Cruz). Cyclin D1 (M-20, Santa Cruz). Cyclin A (C-19, Santa Cruz). Rb (C-15, Santa Cruz). Phospho-Rb (Ser807/811)(9308, Cell Signaling). PCNA (PC10, Santa Cruz). PAX6 (PRB-278P, Covance; Princeton, NJ), p27 (610241, BD-Transduction Laboratories; Franklin Lakes, NJ). CREB (9104, Cell Signaling). Phospho-CREB (Ser133) (9191, Cell Signaling). C-myc (N-262, Santa Cruz). β-actin (AC-15, Sigma-Aldrich). Secondary antibodies goat α-Mouse IgG, DyLightTM 680 and/or goat α-Rabbit Ig-G, DyLightTM 800 (Thermo Fischer Scientific) were used for detection in a Li-Cor Odyssey system (Thermo Fisher Scientific; Waltham, MA).

Moura D.S., Fernández I.F., Marín-Royo G., López-Sánchez I., Martín-Doncel E., Vega F.M, & Lazo P.A. (2016). Oncogenic Sox2 regulates and cooperates with VRK1 in cell cycle progression and differentiation. Scientific Reports, 6, 28532.

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Cellular Protein Quantification and Validation

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Cycloheximide (01810) and calf-intestinal alkaline phosphatase (CIP, P4978) were purchased from Sigma. Dimethyl sulfoxide (D8418, Sigma-Aldrich, St Louis, MO, USA) was added to the cultures at 0.1% (v/v) as a solvent control. Specific antibodies against ATP5C1 (PA5-29975) and NDUFA9 (459100) were purchased from Invitrogen. Antibodies specific for ATP5A1 (51), SIRT3 (14.45), VDAC (B-6), SDHA (B-1), UQCRC2 (G-10), COX IV (D-20), PGC-1α (H-300), PGC-1β (E-9), NRF1 (H-285), NRF2 (C-20), Tfam (H-203), SIRT1 (H-300), PKCα (C-20), actin (I-19), V5 (H-9), Flag (D-8), His (AD1.1.10) and GST (B-14) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Kim Y.R., Kim J.S., Yun J.S., Kim S., Kim S.Y., Jang K, & Yang C.S. (2018). Toxoplasma gondii GRA8 induces ATP5A1–SIRT3-mediated mitochondrial metabolic resuscitation: a potential therapy for sepsis. Experimental & Molecular Medicine, 50(3), e464-.

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