48-8896-42

antibody from Invitrogen Antibodies

Targeting: GZMB CCPI, CGL-1, CGL1, CSP-B, CSPB, CTLA1, CTSGL1, HLP, SECT

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

48-8896-42

Provider

Invitrogen Antibodies

Product name

Granzyme B Monoclonal Antibody (N4TL33), eFluor™ 450, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Applications Reported: This N4TL33 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This N4TL33 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.5 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. eFluor™ 450 is an alternative to Pacific Blue™. eFluor 450 emits at 446 nm and is excited with the violet laser line (405 nm). Please make sure that your instrument is capable of detecting this fluorochrome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

N4TL33

Vial size

100 Tests

Concentration

5 µL/Test

Storage

4° C, store in dark, DO NOT FREEZE!

References

Submitted references

LINC01123 promotes immune escape by sponging miR-214-3p to regulate B7-H3 in head and neck squamous-cell carcinoma.

Li H, Yang Z, Yang X, Zhang F, Wang J, Wu Z, Wanyan C, Meng Q, Gao W, Yang X, Wei J
Cell death & disease 2022 Feb 3;13(2):109

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood cells were stained intracellularly, using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol, with CD8a Monoclonal Antibody, APC (Product # 17-0088-82) and Mouse IgG2a kappa Isotype Control, eFluor 450 (Product # 48-4724-82) (left) or Granzyme B Monoclonal Antibody, eFluor 450 (right). Cells in the lymphocyte gate were used for analysis.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 4 Upregulation of LINC01123 or B7-H3 or downregulation of miR-214-3p induces dysfunction of CD8 + T cells. CD8 + T cells were isolated from PBMCs using immunomagnetic beads. After activation and amplification, CD8 + T cells were cocultured with different CAL-27 cell treatment groups. A Expression of TNF-alpha, IFN-gamma, perforin, and granzyme B in CD8 + T-cell population was analyzed with flow cytometry. B Statistical analysis of the results from flow cytometry of CD8 + T cells. C Expression levels of TNF-alpha, IFN-gamma, perforin, and granzyme B in CAL-27-CD8 + T cocultured supernatants, as detected by ELISA. D CCK8 cytotoxicity test of CD8 + T cells. E ELISA was used to assess the immune activity of CD8 + T cells after recombinant human B7-H3 treatment. * P < 0.05 compared with cells without treatment. The experiment was repeated three times.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 6 The function of LINC01123 in HNSCC can be reversed by miR-214-3p silencing. HNSCC cells were transfected with sh-LINC01123 in the presence of inhibitor-NC or miR-214-3p inhibitor. A Relative expression levels of LINC01123, miR-214-3p, and B7-H3 were determined by qRT-PCR. B B7-H3 protein expression was determined by western blot analysis. C CAL-27 cell viability was measured using a CCK8 assay. D Migration of CAL-27 cells was determined by scratch wound-healing assay. E Statistical results of the scratch-healing test. F Migration and invasion of CAL-27 cells, as determined by transwell migration and invasion assay. G Number of migration and invasion cells. H Apoptosis rate and cell-cycle stage were detected by flow cytometry. I Statistical analysis of apoptosis results. J Statistical analysis of cell cycle stage results. K CD8 + T cells were cocultured with CAL-27 cells that had been transfected with plasmids, and the expression of TNF-alpha, IFN-gamma, perforin, and granzyme B in the CD8 + T cells was analyzed by flow cytometry. L Statistical analysis of CD8 + T-cell flow cytometry results. M After different treatments, CAL-27 cells were cocultured with CD8 + T cells, and the expression levels of TNF-alpha, IFN-gamma, perforin, and granzyme B in coculture supernatants were detected by ELISA. N Killing effect of CD8 + T cells on CAL-27 cells, as assessed by CCK8. O Tumor-volume growth curves of the sh-LINC01123 + inhibitor-NC group and sh-LINC01123 + miR-214-3p-inhibit