Anti-PSMA antibody [EP3253] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PSMA antibody [EP3253] - BSA and Azide free (AB247436)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized LNCaP (human prostate carcinoma epithelial cell) cells labelling PSMA with primary antibody anti-PSMA (ab247436) at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in LNCaP cells. Anti-alpha Tubulin antibody (DM1A) - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue).
Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PSMA antibody [EP3253] - BSA and Azide free (AB247436)

Intracellular Flow Cytometry analysis of LNCaP (Human prostate carcinoma epithelial cell) cells labeling PSMA with ab247436 at 1/100 dilution (1 μg) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti-rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - cell without incubation with primary antibody and secondary antibody (Blue).

• IP

Lab

Immunoprecipitation - Anti-PSMA antibody [EP3253] - BSA and Azide free (AB247436)

PSMA was immunoprecipitated from LNCaP whole cell lysate, 10 µg with ab247436 at 1/50 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab247436 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : LNCaP (human prostate carcinoma epithelial cell) whole cell lysate 10μg. Lane 2 : LNCaP whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab247436 in LNCaP whole cell lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Observed MW : 100 kDa Exposure : 3 seconds

All lanes:

Immunoprecipitation - Anti-PSMA antibody [EP3253] - BSA and Azide free (ab247436) at 1/1000 dilution

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 84 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-PSMA antibody [EP3253] - BSA and Azide free (AB247436)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-PSMA antibody [EP3253] - BSA and Azide free (ab247436) at 1/5000 dilution

Lane 1:

LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method at 20 µg

Lane 2:

LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 84 kDa

Observed band size: 100 kDa

false

Exposure time: 40s

• WB

Lab

Western blot - Anti-PSMA antibody [EP3253] - BSA and Azide free (AB247436)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST Negative control : PC-3 (PMID : 19830782).

All lanes:

Western blot - Anti-PSMA antibody [EP3253] - BSA and Azide free (ab247436) at 1/2000 dilution

Lane 1:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

Lane 2:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method at 20 µg

Lane 3:

LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

Lane 4:

LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/200000 dilution

Predicted band size: 84 kDa

Observed band size: 100 kDa

false

Exposure time: 40s