Human β2m

Manufactured by Merck Group

Sourced in United States

Human β2m is a laboratory-grade reagent used in various immunological and biochemical assays. It is a small protein that is a component of the major histocompatibility complex class I (MHC I) molecule. Human β2m is widely used as a reference standard and control in research and diagnostic applications.

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Lab products found in correlation

Sterile mixed cellulose ester gridded filter paper, by Merck Group (2 mentions) Gelfoam sterile sponge, by Pfizer (2 mentions) Hyclone ccm1 serum free media, by GE Healthcare (2 mentions) Cy5 labeled goat anti human igg h l polyclonal antibody, by Abcam (1 mentions) Pvdf membrane, by Roche (1 mentions) Peroxidase conjugated goat anti mouse secondary antibody, by Jackson ImmunoResearch (1 mentions)

4 protocols using human β2m

Exogenous Peptide Binding Assay

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T2 is a mutant cell line that lacks TAP which allows for efficient loading of exogenous peptides. The 5 × 10⁵ T2 cells were cultured in serum-free IMDM medium containing 25 μg/mL RMF peptide and 5 μg/mL human β2m (Sigma-Aldrich, St. Louis, MO, USA) overnight at 37 °C, 5% CO₂. Then, cells were washed with ice-cold staining buffer (1% BSA in PBS) and incubated with 10 μg/mL ESK or Q2L for 30 min on ice. After staining buffer washing, cells were incubated with secondary Cy5-labeled Goat Anti-Human IgG (H+L) polyclonal antibody (Abcam, Cambridge, UK) and the MFI was measured on ACEA NovoCyteTM flow cytometry.
The 5 × 10⁵ K562-A2-WT1_126–134 cells (Supplementary Materials) or A431 cells were collected and incubated with 10 μg/mL ESK or Q2L and then immunostained and analyzed by flow cytometry as above.

Shen Y., Li Y.M., Zhou J.J., Zhou Z., Xu Y.C., Zhao W.B, & Chen S.Q. (2019). The Antitumor Activity of TCR-Mimic Antibody-Drug Conjugates (TCRm-ADCs) Targeting the Intracellular Wilms Tumor 1 (WT1) Oncoprotein. International Journal of Molecular Sciences, 20(16), 3912.

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Timed Mating and Fetal Thymic Organ Culture

Cited 2 times

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Timed matings were performed, as previously described (58 (link)). Briefly, mouse cages were divided in half using an acrylic divider. One male and one female mouse resided on each side of a divided cage for ∼72 h, after which the divider was removed in late afternoon to allow mating overnight. FTOC was performed as previously described (34 (link)). Fetal thymi were harvested on embryonic day 15. Each fetal thymic lobe was placed on sterile mixed cellulose ester gridded filter paper (Millipore) on top of a Gelfoam sterile sponge (Pfizer), in HyClone CCM1 serum-free media (GE Life Sciences) in one well of a 48-well plate. Some cultures were supplemented with exogenous human β2M (5 μg/mL, Sigma) and peptides FARL (SSIEFARL), Q4H7 (SIIQFEHL), or OVA (SIINFEKL) at the indicated concentrations. Thymi were cultured for 24 h at 37 °C before harvest and analysis by flow cytometry.

Laffey K.G., Stiles R.J., Ludescher M.J., Davis T.R., Khwaja S.S., Bram R.J., Wettstein P.J., Ramachandran V., Parks C.A., Reyes E.E., Ferrer A., Canfield J.M., Johnson C.E., Hammer R.D., Gil D, & Schrum A.G. (2022). Early expression of mature αβ TCR in CD4−CD8− T cell progenitors enables MHC to drive development of T-ALL bearing NOTCH mutations. Proceedings of the National Academy of Sciences of the United States of America, 119(27), e2118529119.

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Fetal Thymic Organ Culture Assay

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Timed matings were performed, as previously described (71 ). Briefly, mouse cages were divided in half using an acrylic divider, with adequate food and water available on each side. One male and one female resided on each side of a divided cage for ~72 hours, after which the divider was removed in late afternoon to make mating possible overnight. FTOC was performed as previously described (35 (link)). On embryonic day (e)15, carbon dioxide was used to euthanize mothers, and fetal thymi were harvested. Each fetal thymic lobe was placed on sterile mixed cellulose ester gridded filter paper (Millipore) on top of a Gelfoam sterile sponge (Pfizer), in HyClone CCM1 serum-free media (GE Life Sciences) in one well of a 48- or 96-well plate. Some cultures were supplemented with exogenous human β2m (5 μg/ml, Sigma) and peptides FARL, Q7, or OVA at the indicated concentrations. Tissue culture occurred at 37°C for 7 days before harvest and assessment of thymic selection by flow cytometry.

Neier S.C., Ferrer A., Wilton K.M., Smith S.E., Kelcher A.M., Pavelko K.D., Canfield J.M., Davis T.R., Stiles R.J., Chen Z., McCluskey J., Burrows S.R., Rossjohn J., Hebrink D.M., Carmona E.M., Limper A.H., Kappes D.J., Wettstein P.J., Johnson A.J., Pease L.R., Daniels M.A., Neuhauser C., Gil D, & Schrum A.G. (2019). The early proximal αβ TCR signalosome specifies thymic selection outcome through a quantitative protein interaction network. Science immunology, 4(32), eaal2201.

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Purity Assessment of CD22c-β2m Protein

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The purity of the CD22c-β2m protein produced was monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with 5 μg of purified CD22c-β2m and human β2m (Sigma-Aldrich, ref M4890) used as a control. After electrophoresis on a 12% acrylamide gel under non-reducing conditions in 1x Tris Glycine SDS running buffer, the proteins were stained using Coomassie brilliant blue (National Diagnostics, ref HS-604) and the gel was scanned using a Bio-Rad scanner.
The SDS-PAGE gel was then transferred onto pore polyvinylidene fluoride (PVDF) membrane (Roche, ref 03010040001) in tris-glycine blotting buffer (Bio-Rad, ref 1610771). Non-specific sites were blocked using a TBS-0.1%-Tween-5% milk buffer, and PVDF membrane was incubated with the primary antibody 6H4 directed against human β2m for 2 h at room temperature. After washing, the membrane was incubated with a peroxidase-conjugated goat anti-mouse secondary antibody (Jackson ImmunoResearch, ref 115-035-003) (1:1,000 dilution) for 90 min. Target proteins were visualized with the Bio-Rad camera after revelation with the 3,3′-Diaminobenzidine (DAB) substrate.

Etienne F., Berthaud M., Nguyen F., Bernardeau K., Maurel C., Bodet-Milin C., Diab M., Abadie J., Gouilleux-Gruart V., Vidal A., Bourgeois M., Chouin N., Ibisch C, & Davodeau F. (2020). SPECT-CT Imaging of Dog Spontaneous Diffuse Large B-Cell Lymphoma Targeting CD22 for the Implementation of a Relevant Preclinical Model for Human. Frontiers in Oncology, 10, 20.

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