MA5-23707

antibody from Invitrogen Antibodies

Targeting: CXCL1 FSP, GRO1, GROa, MGSA, MGSA-a, NAP-3, SCYB1

Western blot (Recommended by provider)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Blocking/Neutralizing (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-23707

Provider

Invitrogen Antibodies

Product name

CXCL1 Monoclonal Antibody (20326)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

In Western blots, this antibody shows approximately 20% cross-reactivity with recombinant human (rh) CXCL2/GRO beta and rhCXCL3/GRO gamma and no cross-reactivity with recombinant rat (rr)CINC-1, rrCINC-2 alpha, rrCINC-2 beta, rrCINC-3, rhMIP-1 alpha, recombinant mouse (rm)MIP-1 alpha, rmMIP-1 beta, rmMIP-1 beta, rhMIP-1 gamma, rmMIP-1 gamma, rmMIP-2, rhMIP-3 alpha, rmMIP-3 alpha, rrMIP-3 alpha, rhMIP-3 beta, or rmMIP-3 beta. Reconstitute at 0.5 mg/mL in sterile PBS. Endoxin level is

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

20326

Vial size

500 µg

Concentration

0.5 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Cancer Burden Is Controlled by Mural Cell-β3-Integrin Regulated Crosstalk with Tumor Cells.

Wong PP, Muñoz-Félix JM, Hijazi M, Kim H, Robinson SD, De Luxán-Delgado B, Rodríguez-Hernández I, Maiques O, Meng YM, Meng Q, Bodrug N, Dukinfield MS, Reynolds LE, Elia G, Clear A, Harwood C, Wang Y, Campbell JJ, Singh R, Zhang P, Schall TJ, Matchett KP, Henderson NC, Szlosarek PW, Dreger SA, Smith S, Jones JL, Gribben JG, Cutillas PR, Meier P, Sanz-Moreno V, Hodivala-Dilke KM
Cell 2020 Jun 11;181(6):1346-1363.e21

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Growth-regulated alpha protein was performed using 70% confluent log phase A-375 cells, treated with10 ng/mL of Human TNF-alpha for 24 hours followed by treatment with protein transport inhibitor cocktail for 4 hours. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CXCL1 Monoclonal Antibody (20326) (Product # MA5-23811) at a concentration of 1 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytosolic localization. Panel e represents A-375 cells treated with protein transport inhibitor cocktail but without TNF-alpha. Panel f represents control A-375 cells with no primary antibody to assess background. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of CXCL1 in Human peripheral blood mononuclear cell (PBMCs) treated with 1 μg/mL LPS for 24 hours and3 μM monensin for 2 hours. Samples were incubated in CXCL1 monoclonal antibody (Product # MA5-23811) 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

Neutralization antibody testing demonstrates the specificity of an antibody through a correlation between antibody binding and the activity of the target. Neutralization of CXCL1 is shown by the decrease in fluorescence (Chemotaxis measured by Resazurin) with increasing concentrations of CXCL1 Antibody (Product # MA5-23707).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Neutralization of CXCL1 in BaF3 mouse pro‚B cell line transfected with human CXCR2. Samples were incubated in CXCL1 monoclonal antibody (Product # MA5-23811). Recombinant Human CXCL1/GROα/KC/CINC‚1 chemoattracts the BaF3 mouse pro‚B cell line transfected with human CXCR2 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin. Chemotaxis elicited by Recombinant Human/Primate CXCL1/GROα/KC/CINC‚1 (10 ng/mL) is neutralized (green line) by increasing concentrations of Human CXCL1/GROα/KC/CINC‚1 Monoclonal Antibody. The ND50 is typically 3-15 µg/mL.