Anti-RPA32/RPA2 antibody [4E4]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 antibody [4E4] (AB252861)

Immunofluorescence analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 (green) with ab252861 at 1/50 dilution, followed by secondary antibody (ab150157) at 1 : 1000 (2μg/mL). Confocal image showing nuclear staining in HeLa cell line.

Counterstain : DAPI (blue) - ab179504 (YCA-R16775) Anti-beta IV Tubulin at 1 : 200 (2.5 μg/mL).

Counterstain Secondary antibody : ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594-Red) at 1 : 500 (20μg/mL).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) secondary.

• I-ELISA

Supplier Data

Indirect ELISA - Anti-RPA32/RPA2 antibody [4E4] (AB252861)

Human RPA32/RPA2= 1μg/mL

ab252861 used at 0-5000 ng/mL.

Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rat IgG (H+L) used at 1 : 1000.

• WB

Supplier Data

Western blot - Anti-RPA32/RPA2 antibody [4E4] (AB252861)

Blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-RPA32/RPA2 antibody [4E4] (ab252861) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate at 20 µg

Lane 2:

HUVEC (human umbilical vein endothelial cell ), whole cell lysate at 20 µg

Lane 3:

U-2 OS (human bone osteosarcoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 29 kDa

false

• WB

Supplier Data

Western blot - Anti-RPA32/RPA2 antibody [4E4] (AB252861)

Blocking buffer : 5% NFDM/TBST

Exposure time : 5 seconds

All lanes:

Western blot - Anti-RPA32/RPA2 antibody [4E4] (ab252861) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 29 kDa

false

• WB

Supplier Data

Western blot - Anti-RPA32/RPA2 antibody [4E4] (AB252861)

Blocking buffer : 5% NFDM/TBST

Exposure time :

lane 1 : 5.5 seconds
lane 2 : 3 seconds

All lanes:

Western blot - Anti-RPA32/RPA2 antibody [4E4] (ab252861) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 29 kDa

false