Cd138 281 2

Manufactured by BioLegend

Sourced in United States

CD138 (281-2) is a mouse monoclonal antibody that recognizes the CD138 antigen, also known as Syndecan-1. CD138 is a transmembrane heparan sulfate proteoglycan that is expressed on plasma cells and a subset of plasmablasts. This antibody can be used for the identification and enumeration of plasma cells by flow cytometry.

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Lab products found in correlation

B220 ra3 6b2, by BioLegend (1 mentions) Cd103 2e7, by BioLegend (1 mentions) Cd68 fa 11, by Bio-Rad (1 mentions) Ly6g 1a8, by BioLegend (1 mentions) Moma 1, by Abcam (1 mentions) Prolong gold mountant, by Thermo Fisher Scientific (1 mentions) Cd5 53 7.3, by Thermo Fisher Scientific (1 mentions) Cd8α 53 6.7, by BioLegend (1 mentions) Cd3 145 2c11, by BioLegend (1 mentions) Cd19 1d3, by BD (1 mentions) Cd4 rm4 5, by BioLegend (1 mentions) B220 ra3 6b2, by BD (1 mentions) Lsrii cytometer, by BD (1 mentions) Biotinylated peanut agglutinin pna, by Vector Laboratories (1 mentions) Lsrfortessa x 20 analyzer, by BD (1 mentions) Cd38 90, by Thermo Fisher Scientific (1 mentions) Anti b220, by BD (1 mentions) Streptavidin bv785, by BioLegend (1 mentions) Anti mouse bcl6 k112 91, by BD (1 mentions) Cd95 jo2, by BD (1 mentions)

Spelling variants of this product

Anti-CD138 (281-2, (4 citations) Anti-CD138-PE-Cy7 (clone 281-2), (3 citations) CD138-BV421 clone 281–2 (2 citations) Usinganti-CD138-APC (clone 281–2, (2 citations) CD138-PE (281–2, (2 citations) APC-CD138 (281-2), (2 citations) Anti-CD138 (clone 281-2, (2 citations)

5 protocols using cd138 281 2

Tissue Sectioning and Immunofluorescence Staining

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Tissue was embedded in optimal cutting temperature medium (TissueTec) and directly snap frozen in isopentane cooled by dry ice. The tissue was then cut into 8- or 14-µm sections by a cryostat and mounted on superfrost glass slides, which were dried for at least 1 h before storage at −70°C. Tissue sections were fixed and permeabilized with ice-cold acetone. The following antibodies were used: B220 (RA3-6B2; BioLegend), CD3 (pc; Abcam), CD68 (FA-11; AbD Serotec), CD103 (2E7; BioLegend), CD138 (281-2; BioLegend), F4/80 (BM8; AbD Serotec), IgG (STAR120F; AbD Serotec), Ly-6G (1A8; BioLegend), MARCO (ED31; AbD Serotec), MOMA1 (Abcam), and PNA (Vector). Tissue was mounted with Prolong Gold mountant (Invitrogen), and images were captured using a confocal scanning microscope (DMI6000; Leica Biosystems) connected to a True Confocal scanner (SP5; Leica Biosystems).

Parsa R., Lund H., Georgoudaki A.M., Zhang X.M., Ortlieb Guerreiro-Cacais A., Grommisch D., Warnecke A., Croxford A.L., Jagodic M., Becher B., Karlsson M.C, & Harris R.A. (2016). BAFF-secreting neutrophils drive plasma cell responses during emergency granulopoiesis. The Journal of Experimental Medicine, 213(8), 1537-1553.

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Multiparameter Flow Cytometry of B, T and CLL Cells

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Peripheral blood mononuclear cells (PBMCs) were blocked for 30 minutes using FBS. Cell surface staining was achieved by incubating cells at 4°C for 30 minutes with the following anti-mouse antibodies: CD3 (145-2C11; Biolegend), IgM (e-Bioscience), B220 (RA3-6B2; BD Pharmingen), CD5 (53-7.3; eBioscience), CD138 (281-2; Biolegend), CD19 (1D3; BD Pharmingen), CD4 (RM4-5; Biolegend) and CD8α (53-6.7; Biolegend). Viability staining was accomplished using DAPI exclusion during acquisition. Apoptotic cells were detected by Annexin V-PE/DAPI staining (BD Pharmingen). Acquisition of B, T and CLL cell populations was performed on a LSRII cytometer (BD Biosciences) harboring a custom configuration for the Wistar Institute. Cytometry data was analyzed using FlowJo software version 7.6.1 (Tree Star Inc.).

Tang C.H., Zundell J.A., Ranatunga S., Lin C., Nefedova Y., Del Valle J.R, & Hu C.C. (2016). Agonist-mediated activation of STING induces apoptosis in malignant B cells. Cancer research, 76(8), 2137-2152.

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Immunohistochemistry for Kidney Tissue Analysis

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Formalin-fixed, paraffin-embedded tissue sections were stained with hematoxylin-eosin (HE), periodic acid-Schiff (PAS) and acid fuchsin orange G (AFOG) according to standard procedures. Specific immunohistochemistry was performed on formalin-fixed, paraffin-embedded consecutive sections using the following antibodies: CD79a (24C2.5; eBioscience, San Diego, CA), CD3 (SP7; NeoMarkers, Freemont, CA), CD138 (281-2; BioLegend, San Diego, CA). Biotinylated peanut agglutinin [PNA] (Ref. B-1075; Vector Laboratories, Burlingame, CA) was used as a marker for germinal center cells. Characterization of infiltrates was performed using polyclonal antibodies against Kappa light chain (Ref. NB7549), Lambda light chain (Ref. NB7552), IgM (Ref. NB7497) and IgA (Ref. NB7504), all Novus Biologicals (Littleton, CO) as well as an anti-polyvalent biotinylated antibody against IgG (Ref. KIT-IDST1007; Empire Genomics, Buffalo, NY). To detect renal IgG deposits, an F(ab’)2 anti-mouse IgG-fluorescein isothiocyanate (FITC) antibody (eBioscience) was used as previously described [65 (link)].

Bauer E., Schlederer M., Scheicher R., Horvath J., Aigner P., Schiefer A.I., Kain R., Regele H., Hoermann G., Steiner G., Kenner L., Sexl V., Villunger A., Moriggl R, & Stoiber D. (2016). Cooperation of ETV6/RUNX1 and BCL2 enhances immunoglobulin production and accelerates glomerulonephritis in transgenic mice. Oncotarget, 7(11), 12191-12205.

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Flow Cytometry Analysis of B Cell Differentiation

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The flow cytometry analysis was performed using the BD LSRFortessa™ X-20 analyzer (BD Biosciences). Mouse splenic cells were stained with anti-B220 (RA3-6B2, BioLegend), CD38 (90, eBioscience), CD95 (Jo2, BD), CD45.1 (A20, BioLegend), and CD45.2 (104, BioLegend) for the in vivo study of B1-8^hi cell differentiation or with anti-B220, CD95, GL7 (GL7, BioLegend), CD138 (281-2, BioLegend), biotin anti-mouse IgG1 (RMG1-1, BioLegend), and BV785 streptavidin (BioLegend) for the in vitro analysis of GC B cell and plasma cell differentiation in the co-culture system or with anti-B220, annexin V (BD), and 7AAD (BD) for the analysis of cell apoptosis. The transduced A20 cell lines (kindly provided by Dr. Jeffrey Ravetch, Rockefeller University) were stained with anti-mouse BCL6 (K112-91, BD) for the knockdown efficiency of Bcl6-specific shRNA.

Zhao R., Zhang H., Zhang Y., Li D., Huang C, & Li F. (2020). In vivo Screen Identifies Zdhhc2 as a Critical Regulator of Germinal Center B Cell Differentiation. Frontiers in Immunology, 11, 1025.

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Comprehensive Immune Profiling of Murine Splenocytes

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Splenocytes were isolated, passed through a 70-µm filter, lysed red blood cells by lysis buffer (ACK buffer: NH4Cl, KHCO3, and EDTA), and placed in staining buffer (PBS + 0.5% BSA). The splenocytes (1 × 10⁶ cells) were stained with following antibody; anti-CD4 (GK1.5), CD8 (53–6.7), CD62l (MEL-14), CD44 (IM7), CD3ε (145–2C11), ICOS (C398.4A), CD11c (N418), B220 (RA3–6B2), CD11b (M1/70), I-Ab (AF6–120.1), PDCA-1 (129c1), CD80 (16–10A1), GL7 (GL7), and CD138 (281–2) (Biolegend, San Diego, CA, USA). The flow cytometry analysis was performed using BDTM LSR-II (BD Biosciences, USA) and FlowJo software (version 10, USA). The serum sample was measured using the LEGENDplex™ Mouse Inflammation Panel kit (IL-1α, IL-1β, IL-6, IL-10, IL-12p70, IL-17A, IL-23, IL-27, MCP-1, IFN-β, IFN-γ, TNF-α, and GM-CSF) (Biolegend, San Diego, CA, USA) and followed the manufacture protocol. The beads were analyzed using BDTM LSR-II (BD Biosciences, USA) and LEGENDplex™ Analysis Software version 8.

Prabakaran T., Troldborg A., Kumpunya S., Alee I., Marinković E., Windross S.J., Nandakumar R., Narita R., Zhang B.C., Carstensen M., Vejvisithsakul P., Marqvorsen M.H., Iversen M.B., Holm C.K., Østergaard L.J., Pedersen F.S., Pisitkun T., Behrendt R., Pisitkun P, & Paludan S.R. (2021). A STING antagonist modulating the interaction with STIM1 blocks ER-to-Golgi trafficking and inhibits lupus pathology. EBioMedicine, 66, 103314.

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