MA5-14470

antibody from Invitrogen Antibodies

Targeting: KLK3 APS, PSA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14470

Provider

Invitrogen Antibodies

Product name

PSA Monoclonal Antibody (EP1588Y)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

MA5-14470 targets Prostate Specific Antigen in IHC (P), ICC/IF and WB applications and shows reactivity with Human samples, however this antibody is not recommended for human colon and mouse prostate tissue sections. This antibody does not react with rat tissue in Western blot applications.

Antibody clone number

EP1588Y

Concentration

Conc. Not Determined

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Prostate Specific Antigen was performed by loading 25 µg of LNCAP (lane 1) and rat prostate (lane 2) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a Prostate Specific Antigen monoclonal antibody (Product # MA5-14470) at a dilution of 1:50 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~29 kDa in LNCAP cells.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Prostate Specific Antigen (green) showing staining in the cytoplasm of PC-3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Prostate Specific Antigen monoclonal antibody (Product # MA5-14470) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Formalin-fixed, paraffin-embedded human prostate stained with rabbit monoclonal PSA using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Prostate Specific Antigen showing positive staining in the Secretion of paraffin-treated Human prostate tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method 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 Prostate Specific Antigen antibody (Product # MA5-14470) diluted by 3% BSA-PBS at a dilution of 1:20-1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively 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.