700656

antibody from Invitrogen Antibodies

Targeting: CXCL5 ENA-78, SCYB5

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

700656

Provider

Invitrogen Antibodies

Product name

CXCL5 Recombinant Rabbit Monoclonal Antibody (9H45L5)

Antibody type

Monoclonal

Antigen

Other

Description

Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Antibody clone number

9H45L5

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CXCL5 was performed by loading 30 µg of A549 (lane1) and A549 treated for 20 min with 200 nM of PMA (lane2) cell lysate using Novex®NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® 2 Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk at 4ºC overnight. CXCL5 was detected at ~ 8 kDa using CXCL5 Recombinant Rabbit Monoclonal Antibody (Product # 700656) at 2-3 µg/mL in 5 % skim milk for 3 hours at room temperature on a rocking platform. Goat anti-Rabbit IgG - HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of CXCL5 showing staining in the cytoplasm of paraffin-embedded human adrenal gland (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a CXCL5 Recombinant Rabbit Monoclonal Antibody (Product # 700656) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

CXCL5 was immunoprecipitated using 5 æg of the CXCL5 Recombinant Rabbit Monoclonal Antibody (Product # 700656) from lysate of A549 cells treated with PMA (200 nM/20 minutes) (Lane 3) using the Dynabeads® Protein A Immunoprecipitation Kit (Product # 10006D). Normal Rabbit IgG was used as a Isotype control (Lane 2). 10 % input represents the cell extract used for immunoprecipitation (Lane 1). Western blot analysis was performed using CXCL5 Recombinant Rabbit Monoclonal Antibody (Product # 700656) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 æg/mL, 1:2500 dilution). Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).