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Np40 ficoll

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NP40-Ficoll is a buffer solution designed for cell lysis and subcellular fractionation. It contains the non-ionic detergent NP40 and the density gradient medium Ficoll, which facilitate the separation and isolation of cellular organelles and macromolecules.

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Lab products found in correlation

Cpg oligonucleotides odn 1826, by InvivoGen (1 mentions) Pneumovax, by Merck Group (1 mentions) Bsa fraction 5, by Merck Group (1 mentions) 96 well elisa plate, by Thermo Fisher Scientific (1 mentions) Pnpp one component substrate, by Southern Biotech (1 mentions) Imject alum, by Thermo Fisher Scientific (1 mentions) Addavax, by Thermo Fisher Scientific (1 mentions)

6 protocols using np40 ficoll

1

Hapten-Carrier Immunization Protocol

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20μg of 4-hydroxy-3-nitrophenylacetatyl hapten conjugated to Ficoll (NP40-Ficoll; BioSearch Technologies) or 100μg of 4-hydroxy-3-nitrophenylacetatyl hapten conjugated to keyhole limpet hemocyanin (NP35-KLH; BioSearch Technologies) was dissolved in 100μL of sterile saline and injected i.p.
Lownik J.C., Wimberly J., Takahashi-Ruiz L, & Martin R.K. (2019). B cell ADAM17 Controls T cell Independent Humoral Immune Responses through Regulation of TACI and CD138. Biochemical and biophysical research communications, 522(2), 442-447.
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2

Immunization with NP-Ficoll and CpG Oligonucleotides

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Mice were immunized intraperitoneally (i.p.) with 50 μg NP40‐Ficoll (Biosearch Technologies, Novato, CA) in 200 μL PBS. CpG oligonucleotides ODN 1826 (InvivoGen) were administered at a dose of 50 μg in 200 μL i.p. LPS of the Salmonella Minnesota strain (Enzo Life Sciences, Farmingdale, NY) was injected intravenously (i.v.) at a dose of 5 μg in 100 μL PBS.
Khoenkhoen S., Erikson E., Ádori M., Stark J.M., Scholz J.L., Cancro M.P., Pedersen G.K, & Karlsson Hedestam G.B. (2019). TACI expression and plasma cell differentiation are impaired in the absence of functional IκBNS. Immunology and Cell Biology, 97(5), 485-497.
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3

NP(40)-Ficoll immunization in mice

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Mice were immunized with 50 µg NP (40)-Ficoll (Biosearch technologies) diluted in PBS and 200 µl was injected intraperitoneally (i.p.).
Pedersen G.K., Li X., Khoenkhoen S., Ádori M., Beutler B, & Karlsson Hedestam G.B. (2018). B-1a Cell Development in Splenectomized Neonatal Mice. Frontiers in Immunology, 9, 1738.
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4

Immunization with NP-ficoll and Pneumovax

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Mice were immunized with 50-μg NP (40)-ficoll (Biosearch Technologies) or a 1:10 dilution of Pneumovax® (Merck & Co.) corresponding to 0.5 μg of each polysaccharide antigen. The antigens were diluted in PBS and 100 μl was injected intraperitoneally (i.p.).
Pedersen G.K., Ádori M., Stark J.M., Khoenkhoen S., Arnold C., Beutler B, & Karlsson Hedestam G.B. (2016). Heterozygous Mutation in IκBNS Leads to Reduced Levels of Natural IgM Antibodies and Impaired Responses to T-Independent Type 2 Antigens. Frontiers in Immunology, 7, 65.
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5

Immunization and Antibody Detection

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Wild-type and mutant 6- to 8-wk-old mice were immunized with either sheep RBCs, NP35-KLH, or NP40-Ficoll. For the sheep RBC immunizations 200 μl of 10% solution of sheep RBCs (Lonza Walkerville, Inc.) was given by intraperitoneal injection. NP35-KLH (Biosearch Technology) was mixed with Imject® Alum (Thermo Scientific) and introduced into mice (100 μg) via intraperitoneal injection. Mice were boosted with same dose of antigen at the indicated days along with Alum. Mice were also immunized with 25 μg NP40-Ficoll (Biosearch Technologies) via intraperitoneal injection. Serum NP specific Ig levels in these mice were analyzed by ELISA. Briefly, 96 well ELISA plates (Nunc) were coated with NP30-BSA (Biosearch Technology) overnight at 4°C, washed and blocked with 1% BSA fraction V (Sigma-Aldrich), serum titers were then added to the plates and incubated 4 h at 4°C. After washing alkaline phosphatase-labeled goat anti-mouse Ig isotype specific antibodies were added for 2 h at room temperature (SouthernBiotech). After washing, PNPP one component substrate (SouthernBiotech) was used to detect the amount of secondary antibody bound.
Boularan C., Hwang I.Y., Kamenyeva O., Park C., Harrison K., Huang Z, & Kehrl J.H. (2015). B-lymphocyte Specific loss of Ric-8A Results in a Gα Protein Deficit and Severe Humoral Immunodeficiency. Journal of immunology (Baltimore, Md. : 1950), 195(5), 2090-2102.
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6

Intraperitoneal and Subcutaneous Mouse Immunization

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Mice were immunized intraperitoneally (i.p.) with 50 μg NP(40)-Ficoll (Biosearch Technologies) or 2x106 IU rSFV-βGal in a total volume of 100 μl PBS. NP(49)-CGG (Biosearch Technologies) was diluted in PBS and mixed 1:1 in AddaVax (Invitrogen). Mice were immunized i.p. with 5 µg or subcutaneously (s.c.) with 25 or 2.5 µg NP(49)-CGG on both flank regions in a volume of 100 μl PBS.
Khoenkhoen S., Ádori M., Solís-Sayago D., Soulier J., Russell J., Beutler B., Pedersen G.K, & Karlsson Hedestam G.B. (2022). IκBNS expression in B cells is dispensable for IgG responses to T cell-dependent antigens. Frontiers in Immunology, 13, 1000755.
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