MA5-23833

antibody from Invitrogen Antibodies

Targeting: CCL19 CKb11, ELC, exodus-3, MIP-3b, SCYA19

Western blot (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-23833

Provider

Invitrogen Antibodies

Product name

MIP-3 beta Monoclonal Antibody (54909)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

54909

Vial size

100 µg

Concentration

1.0 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

CALR-TLR4 Complex Inhibits Non-Small Cell Lung Cancer Progression by Regulating the Migration and Maturation of Dendritic Cells.

Chen R, Huang M, Yang X, Chen XH, Shi MY, Li ZF, Chen ZN, Wang K
Frontiers in oncology 2021;11:743050

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of MIP-3 beta in immersion fixed human peripheral blood mononuclear cells (PBMCs). Samples were incubated in MIP-3 beta monoclonal antibody (Product # MA5-23833) using a dilution of 10 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red) and counterstained with DAPI (blue).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of MIP-3 beta in immersion fixed paraffin-embedded sections of human tonsil. Samples were incubated in MIP-3 beta monoclonal antibody (Product # MA5-23833) using a dilution of 25 µg/mL overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown) and counterstained with hematoxylin (blue).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of MIP-3 beta in Human monocyte-derived dendritic cells. Samples were incubated in MIP-3 beta monoclonal antibody (Product # MA5-23833) or isotype control antibody followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer and permeabilized with Flow Cytometry Permeabilization/Wash Buffer.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 2 mCALR facilitates the migration and maturation of DCs by increasing the secretion of TNFalpha and CCL19. (A) The effect of cell supernatants (NC and mCALR) on DC migration was evaluated by migration assay; ** P < 0.01. (B) The number of invaded DCs in both NC and mCALR groups was quantified by western blot using anti-beta-actin antibody. (C-F) The effect of cell supernatants (NC and mCALR) on DC maturation was determined using flow cytometry with PE-anti-human CD83 antibody; ** P < 0.01, *** P < 0.001. (G-I) The relative mRNA expression levels of CALR, TNFalpha, and CCL19 were detected in NC and mCALR groups using real-time PCR; * P < 0.05, ** P < 0.01, *** P < 0.001. (J, K) The expression levels of TNFalpha and CCL19 were investigated in NC and mCALR groups using western blot; * P < 0.05, ** P < 0.01. (L, M) The expression levels of TNFalpha and CCL19 were investigated in NC and mCALR groups using ELISA; * P < 0.05, *** P < 0.001. (N) The number of invaded DCs in both anti-CCL19+ and anti-CCL19- groups was quantified by western blot using anti-beta-actin antibody. (O-R) The effect of cell supernatants (anti-TNFalpha+ and anti-TNFalpha- groups) on DC maturation was determined using flow cytometry with PE-anti-human CD83 antibody; **P < 0.01.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 mCALR silence inhibits the migration and maturation of DCs by decreasing the secretion of TNFalpha and CCL19. (A) The relative mRNA expression levels of CALR was detected in siCALR and control groups using real-time PCR; *** P < 0.001. (B, C) The effect of cell supernatants (siCtrl and siCALR) on DC migration was evaluated by migration assay; ** P < 0.01, *** P < 0.001. (D-G) The effect of cell supernatants (siCtrl and siCALR) on DC maturation was determined using flow cytometry with PE-anti-human CD83 antibody; ** P < 0.01, *** P < 0.001. (H, I) The relative mRNA expression levels of TNFalpha and CCL19 were detected in siCALR and control groups using real-time PCR; ** P < 0.01, *** P < 0.001. (J, K) The expression levels of CALR, Flag-mCALR, TNFalpha, and CCL19 were investigated in siCALR and control groups using western blot; * P < 0.05, ** P < 0.01, *** P < 0.001.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 mCALR enhances the secretion of TNFalpha and CCL19 via activating CALR-TLR4-MyD88 signaling. (A) The interaction between mCALR and TLR4 was assessed using co-IP assay in NC and mCALR cells, and the IgG antibody was used as a negative control. (B) The relative mRNA expression level of TLR4 was detected in NC and mCALR groups using real-time PCR; ** P < 0.01, *** P < 0.001. (C) The relative mRNA expression level of TLR4 was detected in siCALR and control groups using real-time PCR; * P < 0.05, ** P < 0.01. (D, E) The expression levels of mCALR, TLR4, MyD88, ERK1/2, p-ERK1/2, p38, p-p38, JNK, p-JNK, p65, and p-p65 were investigated in NC and mCALR groups using western blot, and the anti-Flag antibody was used to detect the mCALR protein; ns, P >0.05, * P < 0.05. (F-H) The relative mRNA expression levels of TLR4, TNFalpha, and CCL19 were detected in mCALR cells transfected with different siTLR4 using real-time PCR; ** P < 0.01, *** P < 0.001. (I, J) The expression levels of TLR4, TNFalpha, and CCL19 were investigated in mCALR cells transfected with different siTLR4 using western blot; * P < 0.05, ** P < 0.01.