WB
Western blot analysis of CXCL5 on different lysates with Rabbit anti-CXCL5 antibody at 1/1,000 dilution. Lane 1: A549 cell lysate, Lane 2: A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 16 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of A549 cells starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours labeling CXCL5 with Rabbit anti-CXCL5 antibody at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CXCL5 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours labeling CXCL5. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
CXCL5 was immunoprecipitated from 0.2 mg A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate with Rabbit anti-CXCL5 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-CXCL5 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate (input), Lane 2: Rabbit anti-CXCL5 antibody IP in A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-CXCL5 antibody in A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 38 seconds.| Product Name | CXCL5 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within human CXCL5 aa 65-114 / 114. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIPWB |
| WB:1:1000 ICC/IF:1:500 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CXCL5 |
|---|---|
| Gene Synonyms | SCYB5 ENA-78 |
| Gene Full Name | C-X-C motif chemokine ligand 5 |
| Gene Summary | This gene encodes a protein that is a member of the CXC subfamily of chemokines. Chemokines, which recruit and activate leukocytes, are classified by function (inflammatory or homeostatic) or by structure. This protein is proposed to bind the G-protein coupled receptor chemokine (C-X-C motif) receptor 2 to recruit neutrophils, to promote angiogenesis and to remodel connective tissues. This protein is thought to play a role in cancer cell proliferation, migration, and invasion. [provided by RefSeq, May 2013] |
| Alternative Names | AMCFII antibody C-X-C motif chemokine 5 antibody C-X-C motif chemokine ligand 5 antibody Chemokine (C X C motif) ligand 5 antibody chemokine (C-X-C motif) ligand 5 antibody Cxcl5 antibody CXCL5_HUMAN antibody ENA 78 antibody ENA-78 (8-78) antibody ENA-78(1-78) antibody AMCFII antibody C-X-C motif chemokine 5 antibody C-X-C motif chemokine ligand 5 antibody Chemokine (C X C motif) ligand 5 antibody chemokine (C-X-C motif) ligand 5 antibody Cxcl5 antibody CXCL5_HUMAN antibody ENA 78 antibody ENA-78 (8-78) antibody ENA-78(1-78) antibody ENA-78(9-78) antibody ENA78 antibody Epithelial derived neutrophil activating protein 78 antibody Epithelial-derived neutrophil-activating protein 78 antibody Lipopolysaccharide-induced CXC chemokine antibody Neutrophil activating peptide ENA 78 antibody Neutrophil activating protein 78 antibody Neutrophil-activating peptide ENA-78 antibody neutrophil-activating protein 78 antibody SCYB5 antibody Small inducible cytokine B5 antibody small inducible cytokine subfamily B (Cys-X-Cys) member 5 (epithelial-derived neutrophil-activating peptide 78) antibody small inducible cytokine subfamily B member 5 antibody Small-inducible cytokine B5 antibody |
| Molecular Weight(MW) | 12kDa(Observed band size: 8kDa) |
| Cellular Localization | Secreted. |
WB
Western blot analysis of CXCL5 on different lysates with Rabbit anti-CXCL5 antibody at 1/1,000 dilution. Lane 1: A549 cell lysate, Lane 2: A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 16 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of A549 cells starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours labeling CXCL5 with Rabbit anti-CXCL5 antibody at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CXCL5 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours labeling CXCL5. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
CXCL5 was immunoprecipitated from 0.2 mg A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate with Rabbit anti-CXCL5 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-CXCL5 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate (input), Lane 2: Rabbit anti-CXCL5 antibody IP in A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-CXCL5 antibody in A549 starved overnight treated with 10ng/mL TNF-alpha and 10nM PMA for 24 hours then add 300ng/mL BFA for 3 hours cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 38 seconds.| Application Notes | WB:1:1000 ICC/IF:1:500 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
