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Affi gel columns

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Affi-gel columns are laboratory equipment used for affinity chromatography. They are designed to facilitate the purification and separation of specific molecules, such as proteins, from complex mixtures. The core function of Affi-gel columns is to provide a matrix for the immobilization of ligands, which can then interact with and bind to the target molecules, allowing for their selective isolation and extraction.

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Lab products found in correlation

Flag tag (flag), by Merck Group (2 mentions) Nr1 glun1, by BD (1 mentions) Normal rabbit igg, by Santa Cruz Biotechnology (1 mentions) Synaptophysin, by Merck Group (1 mentions) Glua2 3, by Merck Group (1 mentions) Gephyrin, by Synaptic Systems (1 mentions) Gephyrin, by BD (1 mentions)

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Aurum affi-gel blue mini columns (3 citations)

4 protocols using affi gel columns

1

Antibody Characterization for TMEM163

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The following antibodies were used at the indicated dilutions: rabbit polyclonal antibodies to TMEM163 (1:1000-3000 for WB), P2X3 (Alomone; 1:1000-4000 for WB): mouse monoclonal antibodies to alpha tubulin (Sigma; 1:100,000 for WB), NR1/GluN1 (BD Biosciences; 1:1000 for WB), FLAG (Sigma; 1:1000 for WB): rat monoclonal antibodies to HA (Roche; 1:400 for surface expression). Polyclonal antisera to TMEM163 proteins were raised by injecting rabbits with a GST-TMEM163 fusion protein encoding the first 88 amino acids of TMEM163. Antisera were affinity purified on Affi-gel columns (Bio-Rad) containing the His-tagged TMEM163 fusion proteins.
Salm E.J., Dunn P.J., Shan L., Yamasaki M., Malewicz N.M., Miyazaki T., Park J., Sumioka A., Hamer R.R., He W.W., Morimoto-Tomita M., LaMotte R.H, & Tomita S. (2020). TMEM163 Regulates ATP-Gated P2X Receptor and Behavior. Cell reports, 31(9), 107704.
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2

Purification of Mannose-6-Phosphate Glycoproteins

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4 × 108 cells per culture where flash frozen in liquid nitrogen before to be stored at -80 °C before to be processed by Peter Lobel laboratory at Rutgers University using the protocol described in. Briefly, cellular extracts were loaded on Affigel columns (Biorad) previously coupled with Soluble bovine cation-independent MPR (sCI-MPR). After immobilization of mannose-6-phosphate glycoproteins, the columns were washed prior elution with a solution containing 10 mM free mannose-6phosphate. The eluates were then subjected to mass spectrometry and analyzed using MS-Fit algorithm of the Protein Prospector suite.
Nordlinger A., Del Rio J., Parikh S., Thomas L., Parikh R., Vaknine H., Brenner R., Baschieri F., Robert A, & Khaled M. (2024). Impairing hydrolase transport machinery prevents human melanoma metastasis. Communications Biology, 7, 574.
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3

Antibody Dilutions for Western Blot and Immunostaining

Cited 2 times
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The following antibodies were used with indicated dilutions: rabbit polyclonal antibodies to GABAAR α1 (1:3,000 for IB), γ2 (1:2,000 for IB), α6 (1:1,000 for IB), GluA2/3 (1:1,000 for IB) (Millipore), GABAAR γ2 (1:2,000 for ICC, 1:500 for IHC), Neuroligin-2 (1:3,000 for IB, 1:200 for ICC) (Synaptic systems), rabbit normal IgG (Santa Cruz), Neto2 (0.1 μg/mL for IB) (Zhang et al., 2009 (link)), α2 (1:500 for IB) (Abcam) and α3 (1:500 for IB) (Novus Biologicals): mouse monoclonal antibodies to α1 (1:2,000 for IB), PSD-95 (1:2,000 for IB, 1:1,000 for ICC and IHC) (NeuroMab), β2/3 (1:1,000 for IB), Actin (1:5,000 for IB) (Millipore), NR1 (1:2,000 for IB, 1:500 for IHC), Gephyrin (1:1,000 for IB), GAD65 (1:1,000 for IHC) (BD Biosciences), Gephyrin (1:1,000 for ICC) (Synaptic Systems), Synaptophysin (1:5,000 for IB), Tubulin (1:5,000 for IB), FLAG (1:1,000 for IB) (Sigma), HA (1:1,000 for IB) (Covance): guinea pig polyclonal antibodies to GFP (0.1 μg/mL for IB, ICC and IHC) (Kim et al., 2010 (link)). Polyclonal antisera to Lhfpl4 proteins (GARLH4) were raised by injecting rabbits with a GST-LH4 fusion protein encoding last 52 amino acids of LH4. Antisera were affinity purified on Affi-gel columns (Bio-Rad) containing the His-tagged LH4 fusion proteins (0.1 μg/mL for IB).
Yamasaki T., Hoyos-Ramirez E., Martenson J.S., Morimoto-Tomita M, & Tomita S. (2017). GARLH family proteins stabilize GABAA receptors at synapses. Neuron, 93(5), 1138-1152.e6.
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4

Polyclonal Antibody Generation and Purification

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Polyclonal anti-Msl1 (3208) and anti-Nsl1 (3130) antibodies were generated by immunization of rabbits with the N-terminal (3-210 amino acids) region of mouse Msl1 or C-terminal region (762-1037 amino acids) of mouse Nsl1. The fragment was amplified and cloned in pET28b (Novagen, France) vector to express proteins in E. coli (BL21). For primer sequences see Supplementary file 1. Polyclonal antibodies were purified through Affi-Gel columns (Bio-Rad). For WB analysis anti-Msl1 (3208) or anti-Nsl1 (3130) antibodies were diluted 1:2000.
Ravens S., Fournier M., Ye T., Stierle M., Dembele D., Chavant V, & Tora L. (2014). Mof-associated complexes have overlapping and unique roles in regulating pluripotency in embryonic stem cells and during differentiation. eLife, 3, e02104.
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