Anti-SPON2 antibody [EPR25828A-87] - BSA and Azide free

• ICC

Supplier Data

Immunocytochemistry - Anti-SPON2 antibody [EPR25828A-87] - BSA and Azide free (AB302898)

This data was developed using ab302897, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized LNCaP (human prostate carcinoma epithelial cell) cells labelling SPON2 with ab302897 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing mainly cytoplasmic staining in LNCaP cell line and weak staining in PC-3 cell line. Low expression : PC-3 (PMID : 22615945) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SPON2 antibody [EPR25828A-87] - BSA and Azide free (AB302898)

This data was developed using ab302897, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-3 (human prostate adenocarcinoma epithelial cell, Left) / LNCaP (human prostate carcinoma epithelial cell, Right) cells labelling SPON2 with ab302897 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression : PC-3 (PMID : 22615945).

• I-ELISA

Lab

Indirect ELISA - Anti-SPON2 antibody [EPR25828A-87] - BSA and Azide free (AB302898)

This data was developed using ab302897, the same antibody clone in a different buffer formulation.

Indirect ELISA Human SPON2 Antigen concentration 1000 ng/ml. ab302897 concentration range 1000-0 ng/ml. Secondary antibody Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) used at 1/2500 dilution.

• WB

Supplier Data

Western blot - Anti-SPON2 antibody [EPR25828A-87] - BSA and Azide free (AB302898)

This data was developed using ab302897, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : PC-3, DU 145 (PMID : 22615945 ) Lysate in lane1 was freshly made and used for Western blotting immediately to minimize protein degradation. The band beneath the target band (40kDa) in Lane 2 is likely to be degraded target fragments. Exposure time : Lane 1 : 81 seconds Lane 2-4 : 37 seconds

All lanes:

Western blot - Anti-SPON2 antibody [EPR25828A-87] (<a href='/en-us/products/primary-antibodies/spon2-antibody-epr25828a-87-ab302897'>ab302897</a>) at 1/1000 dilution

Lane 1:

LNCaP (human prostate carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

LNCaP whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

DU145 (human prostate carcinoma epithelial cell), whole cell lysate 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 1/100000 dilution

Observed band size: 40 kDa

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