25-0389-41

antibody from Invitrogen Antibodies

Targeting: CD38

Provider product page for 25-0389-41

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [34]
Comments [0]
Validations
Flow cytometry [2]
Other assay [13]

Submit

Product number: 25-0389-41 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD38 Monoclonal Antibody (HIT2), PE-Cyanine7, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The HIT2 monoclonal antibody reacts with the human CD38 molecule, an approximately 45 kDa type II transmembrane protein. CD38 is expressed by thymocytes, peripheral B cells including plasma cells, activated T cells, and monocytes. CD38 is a counter-receptor for CD31 and has both cyclase and hydrolase enzymatic activity. Applications Reported: This HIT2 antibody has been reported for use in flow cytometric analysis. Applications Tested: This HIT2 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (1 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: HIT2
Vial size: 25 Tests
Concentration: 5 µL/Test
Storage: 4°C, store in dark, DO NOT FREEZE!

CD38 protein structure - 25-0389-41 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4. IFNgamma is required for development of T-bet hi B DN cells and regulates ASC formation and recovery. ( a ) Ingenuity Pathway Analysis (IPA) to identify predicted upstream direct and indirect regulators of the HD B DN Be1 transcriptome. IPA performed using the 427 DEG (B DN Be1 over B DN Be2; FDR < 0.05) identified in the RNA-seq analysis described in Figure 3b . The predicted activation state (z-score of B DN Be1 over B DN Be2) of each regulator/signaling pathway is shown as bar color (orange, activated; blue, inhibited) with predicted upstream regulators sorted in order of significance (overlap P value). Regulators with an overlap P -value

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. Temporally distinct regulation of T-bet hi IRF4 int pre-ASC and ASC development by IFNgamma, R848 and IL-21. Cartoon ( a ) depicting stimulation of CTV-labeled HD B N cells for 3 days with anti-Ig, R848, IL-21 and IFNgamma (Step 1). Cells were washed and re-cultured for 3 days with R848, IFNgamma, and IL-21 (Step 2, +,+ condition) or individual stimuli were included in Step 1 only (+,- condition) or in Step 2 only (-,+ condition). Cells from day 6 cultures containing IFNgamma ( b-e ), R848 ( f-i ) or IL-21 ( j-m ) in Step 1, Step 2 or both steps were analyzed to determine ASC frequencies ( b, f, j ), ASC recovery ( c, g, k ), cell division ( d, h, l ) and total cell recovery ( e, i, m ). Summary of data ( n ) showing that ASC development and recovery from T-bet hi IRF4 int B DN pre-ASCs requires early IFNgamma, R848 and BCR ''priming'' signals and late R848 and IL-21 proliferation and differentiation signals. See Figure 5-figure supplement 1 for representative flow cytometry plots from each culture showing T-bet hi IRF4 int B DN cells on day 3, CD38 hi CD27 + ASCs on day 6 and CTV dilution on day 6. Data are representative of >=3 experiments. The percentage of cells in each division, the frequency of ASCs and cell recovery (total and ASCs) are shown as the mean +-SD of cultures containing purified B N cells from 3 independent healthy donors. All statistical analyses were performed using one-way ANOVA with Tukey''s multiple comparison test. P values *

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6-figure supplement 2. Flow cytometric characterization of B cells activated during the early priming phase in the presence or absence of IFNgamma and IL-2. Day 6 Be.0, Be.IL2, Be.IFNgamma and Be.gamma2 cells were generated as described in Figure 6e . Representative flow cytometry plots from day 6 Be.0, Be.IFNgamma, Be.IL2 and Be.gamma2 cultures showing CD38 hi CD27 + ASCs ( a ) and CTV dilution ( b ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6. IFNgamma cooperates with R848, IL-2 and IL-21 to promote development and recovery of ASCs. ( a-d ) IFNgamma synergizes with subthreshold amounts of TLR7/8 ligand to induce proliferation and differentiation of B N cells. CTV-labeled HD B N cells were activated for 3 days (Step 1) with anti-Ig, IL-2, and increasing concentrations of R848 (as indicated) in the presence or absence of IFNgamma (10 ng/ml). Cells were washed and re-cultured for 3 additional days (Step 2) with IL-21 and the same concentration of R848 that was used in Step 1. B cell division was measured on day 6 in cultures that were activated with IFNgamma (green circles) or without IFNgamma (orange circles) in the presence of no R848 (0 mug/ml, ( a ), high dose R848 (10 mug/ml, ( b ) or low dose R848 (0.1 mug/ml, ( c ). The frequency of CD38 hi CD27 + ASCs ( d ) on day 6 is shown. ( e-i ) IFNgamma cooperates with IL-2 to promote ASC development and recovery. Cartoon ( e ) depicting CTV-labeled HD B N cells activated for 3 days (Step 1) with anti-Ig and R848 alone (Be.0); with anti-Ig +R848+IFNgamma (Be.IFNgamma); with anti-Ig +R848+IL-2 (Be.IL2); or with anti-Ig +R848+IFNgamma+IL-2 (Be.gamma2). Cells were then washed and recultured for an additional 3 days (Step 2) with R848 and IL-21. The percentage of cells that have undergone cell division ( f ), the total cell recovery ( g ), the ASC frequencies ( h ) and total ASCs recovered ( i ) from each day 6 culture are shown. ( j-k ) Early IFNgamma signals regu

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 4 p16 INK4a is a functional barrier to EBV driven proliferation of lymphoblastoid cells. (A) Blueprint of the primary transcript and the spliced mRNA with the three exons of CDKN2A on chromosome 9 encoding the p16 INK4a protein. The target site of the RNP complex within the 1st exon (exon1alpha) (chr9:21,974,678-21,974,827) is shown. (B) Study of the biological effect of the CDKN2A knockout in a time course experiment. WT and p16 KO cells were mixed such that the fraction of the latter was in the order of 10 to 20%, when the cells were infected with WT or DeltaEBNA3C EBV strains. The knockout status of the CDKN2A gene was studied by next generation sequencing to analyze the CD46 locus of the mixed cell populations over time. The fraction of cells with a disabled CDKN2A gene increased in cells infected with DeltaEBNA3C EBV exceeding 80% after eight weeks, whereas the knockout status of CDKN2A in the population of cells infected with WT EBV did not show a clear trend. Results from two biological replicates are shown, additional replicates can be found in S4A Fig . (C) Cell numbers of four different B cell populations were plotted as a function of days post nucleofection (x-axis) versus the format of the cell culture vessel (y-axis) starting with a single well in a 48-well cluster plate. 2x10 6 B cells with an intact CDKN2A locus (WT cells) or cells with an edited CDKN2A gene (p16 KO cells) were infected with wild-type (WT) EBV (left panel) or DeltaEBNA3C EBV (right panel).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Increased exhausted CD8 + T cells in convalescent patients. A. Expression of activation marker CD38 on CD8 + T cells (upper FACS panel). One exemplary dot plot is shown per study group. The bar diagram (lower panel) shows that CD38 expression was significantly higher on CD8 + T cells in convalescent COVID-19 + patients compared with HC. *P-value

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: A schematic overview of our experimental design to generate antigen-specific CD8+ T cells. CD34+ CB cells were expanded for 7 days and CD34+CD38-/low cells were isolated. After 7 days of CD34+ cell expansion, CD34+ cells were enriched using magnetic bead separation. The enriched cells were stained with anti-CD34 and anti-CD38 antibodies to isolate the CD34+CD38-/low HSC population. 81.0% of FSC versus SSC gated cells were CD34+CD38-/low. Gate was determined by the isotype staining control. Data are representative of at least six independent experiments. To induce Notch signaling and early T cell differentiation, CD34+CD38-/low cells were seeded onto DLL1-coated nontissue culture-treated plates. HLA-TCR signaling to direct antigen-specific differentiation was induced using CMVpp65 peptide or epitope-loaded HLA-A*0201 class I tetramers or GIL epitope-loaded HLA-A*0201 class I tetramers. Abbreviations: CM, conditioned medium; CMV, cytomegalovirus; FSC, Forward Scatter; GIL, Influenza M1 virus; HLA, Human leukocyte antigen; SSC, Side Scatter; TCR, T cell receptor.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 7 FIGURE B cells from aged donor mice do not have defects in participating in the GC 10 days after immunization in young recipient mice and fewer B cells from young donor mice are recovered when transferred into aged recipient mice. (a) Representative flow cytometric plots of CD38 - CD95 + GC B cells gated on donor cells from 10-week-old or 93-week-old donor mice in recipient spleens on day 10 post-transfer and immunisation. Numbers adjacent to gates indicate percentage of donor HEL + B220 + cells. Percentage and number of donor-derived GC B cells (Donor HEL + B220 + CD38 - CD95 + ) are plotted on the graphs on the right. (b) Representative flow cytometric plots of IgM + and IgG1 + donor HEL + B cells. Numbers adjacent to gates indicate percentage of donor HEL + B220 + cells. (c, d) Percentage and number of (c) IgM + and (d) IgG1 + B cells derived from donor cells from young or aged mice in recipient spleens 10 days post-transfer and immunisation. (e) Graph showing the percentage and number of IgG1 + GC B cells out of total Donor HEL + B220 + CD38 - CD95 + cells. (f) Schematic diagram of adoptive transfer of B cells from young SW HEL mice into young (8-12 weeks old) or aged (>90 weeks old) mice in which B cells response was analysed 6 days post-transfer and immunisation. (g) Percentage and number of donor HEL + B220 + cells in spleens of young or aged recipient mice 6 days post-transfer and immunisation. (h) Percentage and number of donor-derived GC B cells (Donor HEL + B220

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Morphological and phenotypic features of tumor cells from newly established myeloma PDX. MM.1S cells and mononuclear cells isolated from BM and pleural effusion samples of MM patient were inoculated subcutaneously in NDG mice. When tumors reached 6-8 mm in diameter, tumors were removed and prepared for HE staining, IHC staining, and single-cell suspensions. (A) Representative image of myeloma MM.1S and PDX models. (B) The morphology of tumor cells from myeloma MM.1S and PDX models by Giemsa staining. Scale bar: 5 mum. (C) Representative IHC stainings of tumor tissues. Scale bar: 100 mum. (D) The single-cell suspensions were analyzed by flow cytometry. The doublet or aggregated events were gated out according to side scatter area (SSC-A) and side scatter width (SSC-W). 7-AAD staining was used to gate out dead cells. The expression levels of CD138, CD38, kappa, and lambda were analyzed in mouse CD45 - cells. Myeloma cell line MM.1S was used as a positive myeloma cell control. The left panels of flow charts were isotype controls. PDX, patient-derived xenograft; HE, hematoxylin-eosin; IHC, immunohistochemical.