Anti-Clusterin 抗体 [EPR20729-240] - BSA and Azide free (Capture)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) (AB244626)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Clusterin with ab244626 at 1/100 (4.96 µg/ml) dilution followed by a ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution (Green).

Confocal image showing cytoplasmic staining in HeLa cell line and no staining in Jutkat cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : Jurkat (HPA).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) (AB244626)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Clusterin with ab244626 at 1/2000 (0.496 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The section was incubated with ab244626 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) (AB244626)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CLU KO HeLa (CLU knockout human cervical adenocarcinoma epithelial cell) cells labelling Clusterin with ab244626 at 1/100 (4.96 µg/ml) dilution followed by a ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution (Green).

Confocal image showing cytoplasmic staining in parental HeLa cell line and no staining in CLU KO HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution.

• WB

Lab

Western blot - Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) (AB244626)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

Negative control : Jurkat
In Western blot, ab244626 was shown to bind specifically to Clusterin. Target of interest was observed at 62 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in Clusterin knockout cell line (lane 2).
The expression molecular weight observed is consistent with what has been described in the literature (PMID : 8373947).

All lanes:

Western blot - Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) (ab244626) at 1/1000 dilution

Lane 1:

Wide-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Clusterin knockout HeLa whole cell lysate at 20 µg

Lane 3:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 62 kDa

false

Exposure time: 52s

• sELISA

Unknown

Sandwich ELISA - Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) (AB244626)

Representative standard curve from corresponding SimpleStep ELISA® Kit (ab174447).