Log In Sign up
The largest database of trusted experimental protocols

Affinipure f ab 2 fragment goat anti mouse igm

Manufactured by Jackson ImmunoResearch
Visit Supplier
Sourced in United States

AffiniPure F(ab′)2 fragment goat anti-mouse IgM is a laboratory reagent used for the detection and quantification of mouse IgM in various immunoassays. It is a highly specific and purified antibody fragment prepared from the whole IgG molecule.

Automatically generated - may contain errors

Lab products found in correlation

Catalase, by Beyotime (1 mentions) Ifn α, by Thermo Fisher Scientific (1 mentions) Anti cd40, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) 96 well flat bottom plate, by Corning (1 mentions) Necrostatin 1, by Merck Group (1 mentions) Mouse b cell isolation kit, by Miltenyi Biotec (1 mentions) Mouse ifn α, by Thermo Fisher Scientific (1 mentions) Tlr7 ligand r848, by Enzo Life Sciences (1 mentions) Softmax pro 5, by Molecular Devices (1 mentions) Flipr calcium 4 assay kit, by Molecular Devices (1 mentions) Flexstation 3, by Molecular Devices (1 mentions) Lps 055 b5, by Merck Group (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Mouse vegf quantikine elisa kit, by R&D Systems (1 mentions) Mouse vegf c elisa kit, by Cusabio (1 mentions) Complete protease inhibitor, by Roche (1 mentions)

5 protocols using affinipure f ab 2 fragment goat anti mouse igm

1

Stimulation and Inhibition of Mouse B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human or mouse B cells were cultured in 96-well flat-bottom plates (Corning, Tewksbury, MA, USA) at 5 × 105 cells per well in 200 μl RPMI-1640 (Gibco, Grand Island, NY, USA) supplemented with 10% FBS (Gibco) and antibiotics (penicillin 100 μg/ml, streptomycin 10 μg/ml; Invitrogen Life Sciences, Carlsbad, CA, USA) in a humidified atmosphere of 5% CO2 at 37°C. For stimulation treatment, B cells were divided into four to six groups: Control (grown in normal media), IFN-α (1000 U/ml, eBiosciences, San Diego, CA, USA), R848 (1 μg/ml, Enzo Life Science, Farmingdale, NY, USA), affiniPure F(ab')2 Fragment Goat Anti-Mouse IgM (5 μg/ml, Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA) plus anti-CD40 (2 μg/ml, eBiosciences), two joint stimuli (anti-IgM/CD40+R848) and three joint stimuli (anti-IgM/CD40+R848+IFN-α). Each group of B cells was stimulated respectively on the first day, and the culture medium was changed every other day. Mouse B cells without treatment as the control group were collected immediately after isolation from the spleen, and used to compare the target gene or protein expression level. In addition, mouse B cells were exposed to the stimuli after pretreated with inhibitor zVAD-fmk (Merck, Billerica, MA, USA), Necrostatin-1 (Merck) or Catalase (Beyotime, Nantong, China) for 1 h.
Fan H., Liu F., Dong G., Ren D., Xu Y., Dou J., Wang T., Sun L, & Hou Y. (2014). Activation-induced necroptosis contributes to B-cell lymphopenia in active systemic lupus erythematosus. Cell Death & Disease, 5(9), e1416-.
+ Open protocol
+ Expand
2

Murine B Cell Isolation and Stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lymphocytes of spleen were isolated by ficoll density centrifugation according to standard procedures. B cells were purified using the mouse B cell Isolation Kit (Miltenyi Biotec, USA) and the purity of obtained B cells was always above 92 %. Purified B cells were cultured in RPMI 1640 medium containing 10 % FBS and stimulated with TLR7 ligand R848 (1 μg/ml, Enzo Life Sciences International), TLR9 ligand CpG-1826 (0.3 μM, Invitrogen), AffiniPure F(ab′)2 Fragment Goat Anti-mouse IgM (10 μg/ml, Jackson ImmunoResearch Laboratories) or mouse IFN-α (1000 U/ml, eBioscience).
Dong G., You M., Ding L., Fan H., Liu F., Ren D, & Hou Y. (2015). STING Negatively Regulates Double-Stranded DNA-Activated JAK1-STAT1 Signaling via SHP-1/2 in B Cells. Molecules and Cells, 38(5), 441-451.
+ Open protocol
+ Expand
3

Calcium Flux Assay for Chemokine Response

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded at 105 cells per 100 μl loading medium (RPMI 1640, 10% FBS) into poly-D-lysine coated 96-well black wall, clear-bottom microtiter plates (Nalgene Nunc). An equal volume of assay loading buffer (FLIPR Calcium 4 assay kit, Molecular Devices) in Hanks’ balanced salt solution supplemented with 20 mM HEPES and 2 mM probenecid was added. Cells were incubated for 1 h at 37 °C before adding chemokine or unconjugated AffiniPure F(ab′)2 fragment goat anti-mouse IgM (Jackson ImmunoResearch Laboratories) and then the calcium flux peak was measured using a FlexStation 3 (Molecular Devices). The data was analyzed with SOFT max Pro 5.2 (Molecular Devices). Data is shown as fluorescent counts and the y-axis labeled as Lm1.
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.
+ Open protocol
+ Expand
4

B Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cell proliferation studies were performed using CFSE (Molecular Probes) in a standard dye dilution assay. Purified B cells were stimulated for 96 hours with various combinations of the following reagents: 1 μg/ml CD40 (HM40-3), 1 μg/ml LPS (055:B5, Sigma-Aldrich), recombinant mouse IL-4 (10 ng/ml), or 10 μg/ml AffiniPure F(ab′)2 fragment goat anti-mouse IgM (Jackson ImmunoResearch Laboratories). Data acquisition was done on FACSCanto II flow cytometer. The proliferation index is the average number of cell divisions that a cell in the original population undergoes; the division index is the average number of cell divisions of the responding cells; and the percent cell division is defined as the proliferation index divided by the division index, and multiplying the results by 100, assuming no cell death, were calculated using FlowJo software.
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.
+ Open protocol
+ Expand
5

VEGF Quantification in B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Purified B cells were stimulated with sIgM (AffiniPure F(ab’)2 fragment goat anti-mouse IgM, Jackson ImmunoResearch) with or without additional BAFF (100 ng/ml) and rIL-4 and culture supernatants collected at indicated time point and stored at ≥−20 °C until used for VEGF-A or C protein analysis. In a separate set of experiments, 1 mg of mice mLN were homogenized in 1 ml of lysis buffer supplemented with complete protease inhibitors (Roche) and stored at −80 °C until assayed for VEGFs level using ELISA kits. VEGF-A levels in the culture supernatant or in tissue homogenates were determined using mouse VEGF quantikine ELISA kit (R&D system, Cat No. MMV00), where as VEGF-C levels were determined using mouse VEGF-C ELISA kit (CUSABIO, CSB-E07361m) according to the manufacturer’s instructions. In another set of experiments, mice were sacrificed at indicated time point post infection and intracellular VEGFs expressing B cells were measured as described previously with out any further manipulations36 (link).
Dubey L.K., Karempudi P., Luther S.A., Ludewig B, & Harris N.L. (2017). Interactions between fibroblastic reticular cells and B cells promote mesenteric lymph node lymphangiogenesis. Nature Communications, 8, 367.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!