Log In Sign up
The largest database of trusted experimental protocols

Goat anti mouse igm f ab 2

Manufactured by Jackson ImmunoResearch
Visit Supplier
Sourced in United States

Goat anti-mouse IgM F(ab')2 is a highly specific secondary antibody fragment that recognizes the IgM class of mouse immunoglobulins. It is produced by enzymatic digestion of goat anti-mouse IgM antibodies, resulting in the purification of the F(ab')2 fragment. This antibody fragment can be used in various immunoassays and other applications where the detection of mouse IgM is required.

Automatically generated - may contain errors

Lab products found in correlation

Interleukin 4 (il 4), by Thermo Fisher Scientific (4 mentions) Glutamax, by Thermo Fisher Scientific (4 mentions) Lipopolysaccharides (lps), by Merck Group (3 mentions) Anti cd40, by Thermo Fisher Scientific (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (3 mentions) Goat anti mouse igg f ab 2, by Jackson ImmunoResearch (3 mentions) Dynabeads mouse cd43 untouched b cells, by Thermo Fisher Scientific (2 mentions) Lipopolysaccharides (lps), by Thermo Fisher Scientific (2 mentions) Indo 1 am, by Thermo Fisher Scientific (2 mentions) Ionomycin, by Merck Group (2 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Easysep mouse b cell isolation kit, by STEMCELL (2 mentions) Streptomycin, by Merck Group (2 mentions) Cd43 microbeads, by Thermo Fisher Scientific (2 mentions) Cd38 90 biotin, by Thermo Fisher Scientific (2 mentions) Cd11c n418 biotin, by Thermo Fisher Scientific (2 mentions) Goat anti mouse κ f ab 2, by Southern Biotech (2 mentions) Mouse rbaff, by R&D Systems (2 mentions) Rat anti mouse cd40 1c10, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

Goat anti-mouse IgM (31 citations)

40 protocols using goat anti mouse igm f ab 2

1

Measuring Intracellular Calcium in B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For measurement of relative intracellular free calcium concentration ([Ca2+]i), RBC-depleted splenocytes (1E7/mL in complete medium containing 2% FCS) were stained with anti-B220 and Fab anti-mouse IgG (H+L), while being loaded with 5 μM indo-1 acetoxymethyl ester (INDO-1 AM, Thermofisher) according to the manufacturer’s protocols. Both Indo loading and flow cytometry were performed at room temperature (RT; ~22°C). After being washed once in medium, cells were resuspended at 5E6 cells/mL in RT medium in 500 μL aliquots. Indo-1 was excited with a 355 nm UV laser, and Ca2+-bound indo-1 was detected with a 379/28 bandpass filter; unbound indo-1 was detected with a 524/40 bandpass filter. Induced changes in relative intracellular free calcium concentration ([Ca2+]i) was determined by calculating the ratio of Ca2+ bound/unbound indo-1 signals over time. After data were acquired for 30 s to establish [Ca2+]i baseline, cells were stimulated with the indicated dose of goat F(ab’)2 anti-mouse IgM, rabbit F(ab’)2 anti mouse IgG (H+L), goat F(ab’)2 anti-human IgM Cμ5, or rat anti-mouse IgM [B-7–6] (JacksonImmuno) and data were collected for an additional 2 min 30 s. The relative [Ca2+]i was measured using a Fortessa X-20 flow cytometer and analyzed with FlowJo software.
Wemlinger S.M., Parker Harp C.R., Yu B., Hardy I.R., Seefeldt M., Matsuda J., Mingueneau M., Spilker K.A., Cameron T.O., Larrick J.W., Getahun A, & Cambier J.C. (2022). Preclinical Analysis of Candidate Anti-Human CD79 Therapeutic Antibodies Using a Humanized CD79 Mouse Model. The Journal of Immunology Author Choice, 208(7), 1566-1584.
+ Open protocol
+ Expand
2

In Vitro B Cell Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
MACS-purified (Miltenyi Biotec) CD43− or CD19+ B cells were activated in vitro at a density of 1–3 × 106 cells/ml with 2 μg/ml of anti-CD40 clone HM40-3 (eBiosciences) plus 25 ng/ml of recombinant mouse IL-4 (R&D Systems), 10 μg/ml of goat F(ab′)2 anti-mouse IgM (Jackson Immunoresearch), LPS (20 μg/ml), or LPS + IL-4 (Sigma).
Ha N., Pham D.H., Shahsafaei A., Naruse C., Asano M, & Thai T.H. (2014). HP-1γ Controls High-Affinity Antibody Response to T-Dependent Antigens. Frontiers in Immunology, 5, 271.
+ Open protocol
+ Expand
3

Isolation and Activation of Primary Murine B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary B cells were negatively isolated from 8–12 week old mice using Dynabeads® Mouse CD43 (Untouched B cells) (Life Technologies) per manufacturer’s instructions. Primary B cells were maintained at 106/mL in RPMI (Dibco) supplemented with 10% Fetal Bovine Serum, 50μM β-mercaptoethanol, 100mM sodium pyruvate, 100mM HEPES buffer, and 1% penicillin/streptomycin. B cells were activated with goat f(ab)2 anti- mouse IgM (10μg/ml; Jackson ImmunoResearch), anti-CD40 (1μg/ml; eBioscience) supplemented with IL4 (10ng/ml;), LPS (500ng/ml;), BAFF (100ng/ml; Peprotech) as indicated.
Kong S., Dong H., Song J., Thiruppathi M., Prabhakar B.S., Qiu Q., Lin Z., Chini E., Zhang B, & Fang D. (2015). Deleted in Breast Cancer 1 (DBC1) suppresses B cell activation through RelB and is regulated by IKKα phosphorylation. Journal of immunology (Baltimore, Md. : 1950), 195(8), 3685-3693.
+ Open protocol
+ Expand
4

Murine Lymphocyte Functional Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Thymocytes were prepared as described earlier (de la Fuente et al., 2006 (link)). Splenic T and B cells were purified by negative selection using kits from Miltenyi Biotec. Thymocytes and purified T cells were cultured in medium alone or in wells coated with 2 µg/ml anti-CD3 monoclonal antibody (clone KT3; Abcam) with or without 2 µg/ml anti-CD28 (clone L293; BD) or 40 ng/ml IL-2 (PeproTech). PMA (Sigma-Aldrich) was used at 50 ng/ml, and ionomycin (Sigma-Aldrich) was used at 0.5 µM. Purified B cells were cultured in medium alone or in the presence of goat F(ab′)2 anti–mouse IgM (Jackson ImmunoResearch Laboratories, Inc.), 2 µg/ml anti-CD40 (R&D Systems), or 10 µg/ml LPS (Sigma-Aldrich). 72 h later, the cells were pulsed with 1 µCi [3H]thymidine and counted.
Kumar L., Chou J., Yee C.S., Borzutzky A., Vollmann E.H., von Andrian U.H., Park S.Y., Hollander G., Manis J.P., Poliani P.L, & Geha R.S. (2014). Leucine-rich repeat containing 8A (LRRC8A) is essential for T lymphocyte development and function. The Journal of Experimental Medicine, 211(5), 929-942.
+ Open protocol
+ Expand
5

Measuring Calcium Flux in Murine B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Calcium mobilization was measured using the fluorescent probe Fluo-3-AM. Isolated splenic B cells at 1 × 107 cells/mL were incubated with 10 μ M of Fluo-3-AM (Invitrogen) and 0.002% (v/v) Pluronic F-127 (Invitrogen) for 30 min at 37°C. Cells were then washed and resuspended in supplemented RPMI media for 15 min at 37°C. Cells were then incubated with 10 μg/mL 6G08 (hIgG1 N297Q mutation) or isotype controls for another 15 min at 37°C. Cells were kept warm prior to data acquisition of background fluorescence, followed by the addition of 20 μg/mL goat F(ab’)2 anti-mouse IgM (Jackson). Acquisition was continued for 2.5 min before the addition of 0.6 μ M ionomycin to elicit robust calcium flux.
Simpson A.P., Roghanian A., Oldham R.J., Chan H.T., Penfold C.A., Kim H.J., Inzhelevskaya T., Mockridge C.I., Cox K.L., Bogdanov Y.D., James S., Tutt A.L., Rycroft D., Morley P., Dahal L.N., Teige I., Frendeus B., Beers S.A, & Cragg M.S. (2022). FcγRIIB controls antibody-mediated target cell depletion by ITIM-independent mechanisms. Cell Reports, 40(3), 111099.
+ Open protocol
+ Expand
6

Murine B Cell Activation Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary B cells were negatively isolated from 8–12 week old mice using Dynabeads® Mouse CD43 (Untouched B cells) (Life Technologies) per manufacturer’s instructions. Purified B cells were maintained at 106/mL in RPMI (Dibco) supplemented with 10% Fetal Bovine Serum, 50μM β-mercaptoethanol, 100mM sodium pyruvate, 100mM HEPES buffer, and 1% penicillin/streptomycin. B cells were activated with goat F(ab)2 anti-mouse IgM (10μg/ml; Jackson ImmunoResearch), anti-CD40 (1μg/ml; eBioscience), IL4 (10ng/ml), LPS (500ng/ml), BAFF (100ng/ml; Peprotech) as indicated.
For cell proliferation and immunoglobulin production assays, purified B cells were stained with Cell Trace Carboxyfluorescein diacetatesuccinimidyl ester (CFSE, 5μm; Life Technologies), and cultured at 106 cells/ml for 5 days with indicated stimuli. After 5 days, cells were subjected to flow cytometry and analysis.
Kong S., Thiruppathi M., Qiu Q., Lin Z., Dong H., Chini E.N., Prabhakar B.S, & Fang. D. (2014). Deleted in Breast Cancer 1 (DBC1) is a Suppressor of B cell Activation by Negatively Regulating Alternative NF-κB Transcriptional Activity. Journal of immunology (Baltimore, Md. : 1950), 193(11), 5515-5524.
+ Open protocol
+ Expand
7

B Cell Differentiation Induction

Check if the same lab product or an alternative is used in the 5 most similar protocols
For PC differentiation, B cells of CDLNs were isolated using the mouse CD19 Positive Selection Kit II (STEMCELL, Vancouver, BC, Canada, 18954). Purified B cells were cultured with 10 ng/mL IL-4 (PeproTech, Rocky Hill, NJ, USA), 5 ng/mL IL-5 (PeproTech), 10 μg/mL F(ab′)2 goat anti-mouse IgM (Jackson ImmunoResearch, West Grove, PA, USA) and 80 ng/mL CD40L (PeproTech) in RPMI-1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS), glutamine, 50 μM β-mercaptoethanol, and penicillin/streptomycin with SMI-4a or DMSO vehicle for 4 days at 37 °C.
Li H., Xie L., Zhu L., Li Z., Wang R., Liu X., Huang Z., Chen B., Gao Y., Wei L., He C., Ju R., Liu Y., Liu X., Zheng Y, & Su W. (2022). Multicellular immune dynamics implicate PIM1 as a potential therapeutic target for uveitis. Nature Communications, 13, 5866.
+ Open protocol
+ Expand
8

Insulin Binding in Splenocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Splenocytes (107 cells/ml) were incubated in 37°C RPMI, 2% (wt/wt) BSA and 1 μmol/l Indo-1, AM (Molecular Probes, Eugene, OR, USA) for 30 min. Anti-CD19, Fab anti-IgM and biotinylated insulin were added during Indo loading. Insulin binding was detected by staining with DyLight 650 (Thermo Fisher Scientific) Fab anti-biotin for 15 min. Fab fragments were used to avoid cell stimulation. Cells (2×106/0.4 ml) were analysed using an LSR Fortessa X-20 flow cytometer to establish a baseline, and then stimulated with 5μg F(ab’)2 goat anti-mouse IgM (Jackson Labs).
Smith M.J., Hinman R.M., Getahun A., Kim S., Packard T.A, & Cambier J.C. (2018). Silencing of high-affinity insulin-reactive B lymphocytes by anergy and impact of the NOD genetic background in mice. Diabetologia, 61(12), 2621-2632.
+ Open protocol
+ Expand
9

Intracellular Ca2+ Measurement in B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
B cells were loaded for 60 min at 37°C with Indo-1 acetoxymethyl (final concentration, 1 µM; Molecular Probes) in RPMI containing 10% FCS. Baseline measurements were acquired for 30 s, and then cells were stimulated with 10 µg/ml F(ab′)2 goat-anti-mouse IgM (Jackson ImmunoResearch) or 2 µg/ml TG2 for 2.5 min. Mean relative [Ca2+]i was monitored over time using an LSR II flow cytometer (BD) with analysis using FlowJo software (Tree Star).
du Pré M.F., Blazevski J., Dewan A.E., Stamnaes J., Kanduri C., Sandve G.K., Johannesen M.K., Lindstad C.B., Hnida K., Fugger L., Melino G., Qiao S.W, & Sollid L.M. (2019). B cell tolerance and antibody production to the celiac disease autoantigen transglutaminase 2. The Journal of Experimental Medicine, 217(2), e20190860.
+ Open protocol
+ Expand
10

Splenic B Cell Activation and IgA Class Switching

Check if the same lab product or an alternative is used in the 5 most similar protocols
For CCR6 upregulation, splenic B cells were stimulated with 10 ug/ml anti-IgM (F(ab’)2 goat anti-mouse IgM, Jackson Immunoresearch) for the indicated time. For IgA class switch B-DC coculture experiments, MACS-isolated splenic B cells (typically at 50,000 cells per well) were stimulated with 10 ug/ml anti-IgM and 20 ug/ml anti-CD40 (clone FGK4.5, UCSF Hybridoma Core) in the presence or absence of sorted DCs at a ratio of 1:1 for 5 days. The sorted DCs were from PPs of untreated mice in all cases except for the experiment involving sorted DC subsets where they were from B16-Flt3L treated mice. For IgA class switch in the absence of DCs, MACS-isolated splenic B cells were stimulated with 10 ug/ml anti-IgM and 20 ug/ml anti-CD40 (clone FGK4.5, UCSF Hybridoma Core) in the presence of TGFβ (2 ng/ml) and RA (100 nM) for 5 days.
For BMDCs, 5 × 106 BM cells were cultured in 10 cm tissue culture dishes in 10 ml of medium supplemented with supernatants from 3T3 cells transfected with the gene-encoding murine GM-CSF 7 days. Cells were treated with 1 μg/ml LTβR agonistic antibody (clone 3C8) and 100 nM RA every 3.5 days for 7 days.
Reboldi A., Arnon T.I., Rodda L.B., Atakilit A., Sheppard D, & Cyster J.G. (2016). IgA production requires B cell interaction with subepithelial dendritic cells in Peyer’s patches. Science (New York, N.Y.), 352(6287), aaf4822.
+ 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!