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Bax n20 antibody

Manufactured by Santa Cruz Biotechnology
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The BAX N20 antibody is a rabbit polyclonal antibody that recognizes the N-terminal region of the BAX protein. BAX is a pro-apoptotic member of the Bcl-2 family of proteins that is involved in the regulation of apoptosis. The BAX N20 antibody can be used to detect and analyze the expression of BAX in various biological samples.

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Lab products found in correlation

Bca kit, by Thermo Fisher Scientific (3 mentions) Bax 6a7 antibody, by Santa Cruz Biotechnology (2 mentions) Protein g mag sepharose, by GE Healthcare (1 mentions) Streptavidin agarose bead, by Thermo Fisher Scientific (1 mentions) Hela cell, by American Type Culture Collection (1 mentions) Protein a g bead, by Thermo Fisher Scientific (1 mentions) Protein a g agarose beads, by Thermo Fisher Scientific (1 mentions) X tremegene, by Roche (1 mentions)

Close spelling variants of this product

Bax antibody (18 citations) Bax (N20) (9 citations)

4 protocols using bax n20 antibody

1

Bax Protein Immunoprecipitation from A549 Cells and Lung Tissue

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Homogenized A549 cells and mice lung tissues were incubated with Bax 6A7 antibody (Santa Cruz Biotechnology) for 6 h and mixed using rotary mixer NRC-200 (NISSEN, Tokyo, Japan) at 4°C temperature. Next, samples were incubated with protein G Mag Sepharose (GE Healthcare UK Ltd, Little Chalfont, Buckinghamshire, UK) for 6 h again. The antibody/antigen complex was pulled out of the sample using magnetic interaction between protein G Mag and Magrock6 (GE Healthcare UK Ltd). This isolated protein was separated by SDS-PAGE for western blot analysis. The Bax protein was detected using Bax N-20 antibody (Santa Cruz Biotechnology) after electrophoresis and transfer.
Suzuki K., Yanagihara T., Yokoyama T., Maeyama T., Ogata-Suetsugu S., Arimura-Omori M., Mikumo H., Hamada N., Harada E., Kuwano K., Harada T, & Nakanishi Y. (2017). Bax-inhibiting peptide attenuates bleomycin-induced lung injury in mice. Biology Open, 6(12), 1869-1875.
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2

Affinity Pulldown of BAX in HeLa Cells

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HeLa cells (ATCC) were cultured in DMEM with 10% FBS and penicillin/streptomycin. To generate whole-cell lysates, HeLa cells (6.0 × 106) were trypsinized, washed with cold PBS, and lysed by incubation with 0.5% CHAPS in lysis buffer (150 mM NaCl, 50 mM Tris [pH 7.4]). The total protein concentration of the soluble fraction was measured using a BCA kit according to the manufacturer’s instructions (Thermo Scientific) and diluted to 1 mg/ml. Biotinylated SAHB (20 μM) was added to 500 μl of pre-cleared lysate and incubated for 16 hr at 4°C. The samples were subjected to affinity pull-down by incubation with streptavidin agarose beads (Thermo Scientific), which were successively washed with ice-cold PBS. Following elution into LDS sample buffer, samples were analyzed by western blotting using the BAX N20 antibody (Santa Cruz).
Barclay L.A., Wales T.E., Garner T.P., Wachter F., Lee S., Guerra R.M., Stewart M.L., Braun C.R., Bird G.H., Gavathiotis E., Engen J.R, & Walensky L.D. (2015). Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism. Molecular cell, 57(5), 873-886.
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3

Regulation of BAX Activation by BCL-2 BH4 Peptides

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Recombinant BAX (1 μM) was mixed with 8 μl liposomes,1 μM BIM SAHBA (aa 145–164), and the indicated doses of BCL-2 BH4 SAHBA or BCL-2 BH4 SAHBA L23A in 20 μl liposomal buffer (10 mM HEPES [pH 7.0], 200 mM KCl, 1 mM MgCl2). Following the indicated incubation period at room temperature, the mixture was added to 280 μl 3% BSA in PBS, and 30 μl of the resulting mixture (10%) was reserved for input analysis. Activated BAX was immunoprecipitated with BAX 6A7 antibody (Santa Cruz Biotechnology) conjugated to pre-washed protein A/G beads (Thermo Scientific) or beads alone, and resolved by electrophoresis. Immunoblotting of the inputs and immunoprecipitated samples was performed with the BAX N20 antibody (Santa Cruz).
Barclay L.A., Wales T.E., Garner T.P., Wachter F., Lee S., Guerra R.M., Stewart M.L., Braun C.R., Bird G.H., Gavathiotis E., Engen J.R, & Walensky L.D. (2015). Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism. Molecular cell, 57(5), 873-886.
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4

Analyzing Bax-Bcl-2 Protein Interactions

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Bax and Bcl-2 cDNAs were subcloned into the pBI-CMV1 bidirectional expression vector (Clontech Laboratories), and the L23A Bcl-2 point mutation was generated by site-directed mutagenesis and confirmed by DNA sequencing. The indicated plasmids were transfected with Xtreme-Gene (Roche) according to the manufacturer’s instructions into HeLa cells (2.5 × 105) that were plated in 6-well format 24 hr prior to transfection. The cells were lysed 24 hr post-transfection in buffer containing 0.5% NP-40 and pre-cleared with protein A/G agarose beads (Thermo Scientific). Pre-clarified lysates were immunoprecipitated with BCL-2 antibody (100) (Santa Cruz). Following three successive washes with cold lysis buffer, immunoprecipitated protein was eluted into LDS buffer and analyzed by western blotting using the BAX N20 antibody (Santa Cruz).
Barclay L.A., Wales T.E., Garner T.P., Wachter F., Lee S., Guerra R.M., Stewart M.L., Braun C.R., Bird G.H., Gavathiotis E., Engen J.R, & Walensky L.D. (2015). Inhibition of Pro-Apoptotic BAX by a Noncanonical Interaction Mechanism. Molecular cell, 57(5), 873-886.
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