Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed)

• WB

Collaborator

Western blot - Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (AB32020)

Blocking and diluting buffer and concentration : 5% Milk + PBST.

In Western blot, β-actin was used as loading control staining at 1/1000 dilution.

Approx. 75k cells of total protein were loaded to all wells.

This product is only suitable for use with overexpressed samples. Additional testing has now shown that in endogenous samples a non-specific band is observed at the same MW as hTERT (120 kDa). We do not know the identity of this band.

Non-Specific band observed is consistent with what has been described in the literature (PMID : 24990373).

All lanes:

Western blot - Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (ab32020) at 1/500 dilution

Lane 1:

NCI-H1299 (human lung carcinoma epithelial cell) Knock Out whole cell lysate at 75000 Cells

Lane 2:

NCI-H1299 transfected with a human TERT expression vector, whole cell lysate at 75000 Cells

Lane 3:

NCI-H1299 whole cell lysate at 75000 Cells

Secondary

Lanes 1 - 3:

Anti-Rabbit for TERT detection

Lanes 1 - 3:

Anti-Mouse for β-actin detection

Observed band size: 120 kDa

false

The data was kindly provided by an Anonymous collaborator

• WB

Collaborator

Western blot - Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (AB32020)

Blocking and diluting buffer and concentration : 5% Milk + PBST.

In Western blot, β-actin was used as loading control staining at 1/1000 dilution.

Approx. 75k cells of total protein were loaded to all wells.

This product is only suitable for use with overexpressed samples. Additional testing has now shown that in endogenous samples a non-specific band is observed at the same MW as hTERT (120 kDa). We do not know the identity of this band.

Non-Specific band observed is consistent with what has been described in the literature (PMID : 24990373).

All lanes:

Western blot - Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (ab32020) at 1/500 dilution

Lane 1:

Calu-6 (Anaplastic carcinoma epithelial cell) transfected with a human TERT expression vector at 75000 Cells

Lane 2:

Calu-6 whole cell lysate at 75000 Cells

Lane 3:

Calu-6 whole cell lysate Knock Down with Antisense oligonucleotides at 75000 Cells

Secondary

Lanes 1 - 3:

Anti-Rabbit for TERT detection

Lanes 1 - 3:

Anti-Mouse for β-actin detection

Observed band size: 120 kDa

false

The data was kindly provided by an Anonymous collaborator

• WB

PubMed

Western blot - Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (AB32020)

Overexpression of either hTR or hTERT increases total telomerase activity in living cells.

(a) hTR or hTERT overexpression in HEK 293T and HeLa cells was quantified by northern or western blot, respectively, with H1 RNA or β-actin as loading controls.

(b) Quantification of telomerase activity as measured by direct telomerase assay using lysates from HEK 293T or HeLa cells untransfected or transfected with either hTR or hTERT expression vectors (error bars : SD, n = 3).

All lanes:

Western blot - Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (ab32020) at 1/1000 dilution

All lanes:

hTR or hTERT overexpression in HEK 293T and HeLa cells

false

Linghe Xi et al Nucleic Acids Res. 2014 Jul 2;42(13):8565–8577. Fig 4. doi: 10.1093/nar/gku560. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/3.0/

• WB

Lab

Western blot - Anti-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (AB32020)

Rabbit Monoclonal [Y182] to Telomerase reverse transcriptase ab32020 staining at 1/1000 dilution, shown in green

Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 127 kDa in TERT HEK293T Overexpression Lysate (Flag tag) cell lysates with no signal observed at this size in HEK293T Mock Transfection Lysate 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 (TBST) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBST, 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-Telomerase reverse transcriptase antibody [Y182] (Overexpressed) (ab32020) at 1/1000 dilution

Lane 1:

TERT HEK-293T Overexpression Lysate (Flag tag) at 20 µg

Lane 2:

HEK-293T Mock Transfection Lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 127 kDa

false