Anti-Clusterin antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody (AB69644)

ab69644 at 1/50 dilution staining Apolipoprotein J in human brain by Immunohistochemistry, Paraffin-embedded tissue, in the absence or presence of the immunising peptide.

• WB

Unknown

Western blot - Anti-Clusterin antibody (AB69644)

All lanes:

Western blot - Anti-Clusterin antibody (ab69644) at 1/500 dilution

Lane 1:

HeLa cell extracts at 30 µg

Lane 2:

A549 extracts at 30 µg

Lane 3:

A549 extracts at 30 µg with immunising peptide

Predicted band size: 52 kDa

Observed band size: 28 kDa,52 kDa

false

• WB

Lab

Western blot - Anti-Clusterin antibody (AB69644)

Western blot : Rabbit Polyclonal to Clusterin ab69644 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in CLU knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Clusterin antibody (ab69644) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

Human CLU knockout U-87 MG cell line at 20 µg

Lane 3:

Human testis at 20 µg

Lane 4:

Ramos at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 70 kDa

false

• WB

Supplier Data

Western blot - Anti-Clusterin antibody (AB69644)

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

Negative control : Jurkat

In Western blot, ab323182 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).

Recombinant multiclonal ab323182 performs better than polyclonal in WB application.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Recombinant multiclonal ab323182 : 114 seconds; Polyclonal ab69644 : 180 seconds

All lanes:

Western blot - Anti-Clusterin antibody [RM1292] (<a href='/en-us/products/primary-antibodies/clusterin-antibody-rm1292-ab323182'>ab323182</a>) 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:

Huh7 (human hepatocelullar carcinoma 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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 62 kDa,36 kDa

false

• WB

CiteAb

Western blot - Anti-Clusterin antibody (AB69644)

Clusterin western blot using anti-Clusterin antibody ab69644. Publication image and figure legend from Lin, Y. M., Sun, H. Y., et al., 2018, Viruses, PubMed 29385741.

ab69644 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab69644 please see the product overview.

Effects of calcitriol on apoJ and Grp78 in HCV infection. HCV-infected Huh7.5-SEAP cells (7 × 105) were pretreated with 5 μM of Wy14643, 10 μM of linoleic acid, 30 μM of Ly171883, or 20 μM of kifunensine for 1 h before calcitriol treatment in a 6-well plate. Cell lysates were collected at day 6 and the apo J, Grp78 and β-actin levels were detected by Western blot. Data are expressed as mean ± SD obtained from three individual cultures. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the medium control cells without HCV. #p < 0.05 and ##p < 0.01 vs. the HCV-infected cells. &p < 0.05 and &&p < 0.01 vs. the calcitriol-treated group. The symbol (–) and (+) referred to the cells without and with HCV infection, respectively.

false