Proteina g plus agarose bead slurry

Manufactured by Santa Cruz Biotechnology

Sourced in United States

ProteinA/G plus–agarose bead slurry is a laboratory product that consists of Protein A and Protein G coupled to agarose beads suspended in a buffer solution. Protein A and Protein G are bacterial proteins that have high affinity for the Fc region of immunoglobulins, making this product useful for the purification and isolation of antibodies and antibody-containing samples.

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Lab products found in correlation

Horseradish peroxidaseconjugates, by Santa Cruz Biotechnology (2 mentions) Ofmonoclonal antibody against gfp, by Santa Cruz Biotechnology (2 mentions) Withantibody horseradish peroxidase conjugates, by Santa Cruz Biotechnology (2 mentions) Laemmli buffer, by Bio-Rad (1 mentions) Amersham imager, by Cytiva (1 mentions) Amersham ecl prime, by Cytiva (1 mentions) Atp11a, by Abcam (1 mentions)

Spelling variants of this product

Agarose beads (51 citations) Agarose (14 citations) G Plus agarose beads (4 citations)

5 protocols using proteina g plus agarose bead slurry

GFP-Foxc2 Interaction Analysis

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Cell lysates (1000 μg of protein) from cells
transfected with GFP-Foxc2 were incubated with 10 μl of
monoclonal antibody against GFP (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at
4°C overnight with of 20 μl of 50% (v/v) Protein
A/G plus–agarose bead slurry (Santa Cruz Biotechnology, Santa Cruz, CA, USA).
Beads were pelleted by centrifugation for 2 min at 1000 g, and then
washed three times with lysis buffer, and twice with PBS. Proteins were eluted and
prepared for analysis by heating in reduced Laemmli sample buffer at 100°C for 5
min. Proteins were resolved by 4–15% gradient SDS/PAGE and transferred on
to PVDF membranes for Western blotting. Membranes were blocked with 5% (w/v)
non-fat dried milk in Tris-buffered saline, then probed either with
antibody–horseradish peroxidase conjugates containing antibody against GFP or CK2
proteins (Santa Cruz Biotechnology, Santa Cruz, CA, USA), then horseradish peroxidase
conjugates (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Golden D, & Cantley L.G. (2014). Casein Kinase 2 prevents mesenchymal transformation by maintaining Foxc2 in the cytoplasm. Oncogene, 34(36), 4702-4712.

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Foxp3 Expression in iTregs with GANT61

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Naïve CD4⁺ T cells were isolated from spleens of naïve mice and differentiated into iTregs and treated +/− 20μM GANT61 or Veh for 24 hours. Viability was assessed for all groups and determined to be greater than 85%. 1 mg of protein lysate was pre-cleared using Protein A/G Plus Agarose bead slurry (Santa Cruz) for 1 hour at 4°C. Pre-cleared lysates were incubated with 2μg RL2 antibody for 3 hours at 4°C. Agarose bead slurry was added and mixed at 4°C overnight. After overnight incubation, beads were spun at 3000 RPM 5 minutes and supernatant was discarded. Beads were washed twice in RIPA buffer then washed 3 times in PBS 5 minutes each at 4°C. Beads were resuspended in 25ul 2X Laemmli Buffer (Bio-Rad) and heated at 95°C; supernatant was collected and resolved on SDS PAGE gel. Blots were probed for Foxp3 or total STAT3. GE Amersham ECL prime western blotting detection reagent (Fisher Scientific) was used for development following manufacturers protocol, and imaged using a Amersham Imager 600 chemiluminescence imaging system.

Hinshaw D.C., Benavides G.A., Metge B.J., Swain C.A., Kammerud S.C., Alsheikh H.A., Elhamamsy A., Chen D., Darley-Usmar V., Rathmell J.C., Welner R.S., Samant R.S, & Shevde L.A. (2023). Hedgehog signaling regulates Treg to Th17 conversion through metabolic rewiring in breast cancer. Cancer immunology research, 11(5), 687-702.

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Immunoprecipitation of O-GlcNAcylated Proteins

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RAW 264.7 cells were polarized using 20ng/ml IL-4 and 20ng/ml M-CSF with 20μM GANT61 or DMSO for 24 hours. 1 mg of protein lysate was pre-cleared using Protein A/G Plus Agarose bead slurry (50μl; Santa Cruz) for 1 hour at 4°C. Pre-cleared lysates were incubated with 2μg RL2 antibody for 2 hours at 4°C and 50μl of Protein A/G Plus Agarose bead slurry was added at 4°C overnight. Beads were resuspended in 2X Laemmli Buffer and heated at 95°C; supernatant was resolved on SDS PAGE gel.

Hinshaw D.C., Hanna A., Lama-Sherpa T., Metge B., Kammerud S.C., Benavides G.A., Kumar A., Alsheikh H.A., Mota M., Chen D., Ballinger S.W., Rathmell J.C., Ponnazhagan S., Darley-Usmar V., Samant R.S, & Shevde L.A. (2021). Hedgehog signaling regulates metabolism and polarization of mammary tumor-associated macrophages. Cancer research, 81(21), 5425-5437.

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ATP11A Protein Immunoprecipitation

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For immunoprecipitation, Santa Cruz Biotechnologys Immunoprecipitation protocol was used. Entire protein lysates were extracted using RIPA lysis buffer supplemented with 1 mM phenylmethylsulfonyl fluoride (PMSF), protease inhibitor, phosphatase inhibitor (Bimake, Houston, TX, USA). Lysates were precleared by adding a 1:2 protein A/G PLUS agarose bead slurry (Santa Cruz Biotechnology, Santa Cruz, CA, USA), incubating for 10 min with agitation, centrifuging, and retaining the resulting supernatant. The rabbit polyclonal antibody ATP11A (Abcam, Cambridge, UK) and mouse monoclonal antibody IgG (Santa Cruz Biotechnology, Santa Cruz, CA, USA) antibodies were preincubated with magnetic beads. Next, the antibody-beads complex was washed three times with lysis buffer and incubated with the soluble supernatants of the protein lysates overnight at 4 °C. After centrifuging the samples at 14,000 rpm, the pellet was washed with PBS, resuspended, and boiled for 5 min to remove immunocomplexes from the beads before centrifugation to pellet the beads. Subsequently, the material was eluted and performed a SDS-PAGE test and western blot analysis.

Chen L., Tang J., Sheng W., Sun J., Ma Y, & Dong M. (2022). ATP11A promotes EMT by regulating Numb PRRL in pancreatic cancer cells. PeerJ, 10, e13172.

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GFP-Foxc2 Interaction Analysis

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Golden D, & Cantley L.G. (2014). Casein Kinase 2 prevents mesenchymal transformation by maintaining Foxc2 in the cytoplasm. Oncogene, 34(36), 4702-4712.

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