Anti-Cathepsin L + V antibody [33/2]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L + V antibody [33/2] (AB6314)

IHC image of Cathepsin L staining in human colon adenocarcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab6314, 0.5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• WB

Project

Western blot - Anti-Cathepsin L + V antibody [33/2] (AB6314)

Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/1000 dilution

Lane 1:

Lung (Human) Tissue Lysate at 20 µg

Lane 2:

Lung (Mouse) Tissue Lysate at 20 µg

Lane 3:

Human kidney tissue lysate - total protein (<a href='/en-us/products/unavailable/human-kidney-tissue-lysate-total-protein-ab30203'>ab30203</a>) at 20 µg

Lane 4:

Kidney (Mouse) Tissue Lysate at 20 µg

Lane 5:

A549 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 38 kDa

Observed band size: 25 kDa,55 kDa

true

Exposure time: 20min

• WB

AbReview24674****

Western blot - Anti-Cathepsin L + V antibody [33/2] (AB6314)

All lanes:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/750 dilution

All lanes:

whole cell lysate prepared from U87MG human glioblastoma at 50 µg

Secondary

All lanes:

Goat anti-mouse HRP at 1/2000 dilution

Predicted band size: 38 kDa

false

Image courtesy of Ratnakar Singh by Abreview.

• WB

Supplier Data

Western blot - Anti-Cathepsin L + V antibody [33/2] (AB6314)

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM/TBST

Observed MW : 30

Exposure time : 3 minutes

Lane 1:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/20 dilution

Lane 1:

Western blot - Anti-Cathepsin L/V/K/H antibody [EPR8011] (<a href='/en-us/products/primary-antibodies/cathepsin-l-v-k-h-antibody-epr8011-ab133641'>ab133641</a>) at 1/20 dilution

Lane 2:

Western blot - Anti-Cathepsin L/V/K/H antibody [EPR8011] (<a href='/en-us/products/primary-antibodies/cathepsin-l-v-k-h-antibody-epr8011-ab133641'>ab133641</a>) at 1/100 dilution

Lane 2:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/100 dilution

Lane 3:

Anti-His tag at 1/10000 dilution

All lanes:

Human Cathepsin L recombinant protein fraction

Secondary

Lanes 1 and 3:

Peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/1000 dilution

Lane 2:

Rabbit anti-Mouse IgG at 1/2000 dilution

Predicted band size: 38 kDa

false

• WB

Supplier Data

Western blot - Anti-Cathepsin L + V antibody [33/2] (AB6314)

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM/TBST

Observed MW : 30

Exposure time : 3 minutes

Lane 1:

Western blot - Anti-Cathepsin L/V/K/H antibody [EPR8011] (<a href='/en-us/products/primary-antibodies/cathepsin-l-v-k-h-antibody-epr8011-ab133641'>ab133641</a>) at 1/20 dilution

Lane 1:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/20 dilution

Lanes 2 and 4:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/100 dilution

Lanes 2 and 4:

Western blot - Anti-Cathepsin L/V/K/H antibody [EPR8011] (<a href='/en-us/products/primary-antibodies/cathepsin-l-v-k-h-antibody-epr8011-ab133641'>ab133641</a>) at 1/100 dilution

Lane 3:

Western blot - Anti-Cathepsin L/V/K/H antibody [EPR8011] (<a href='/en-us/products/primary-antibodies/cathepsin-l-v-k-h-antibody-epr8011-ab133641'>ab133641</a>) at 1/500 dilution

Lane 3:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/500 dilution

Lane 5:

Western blot - Anti-Cathepsin L/V/K/H antibody [EPR8011] (<a href='/en-us/products/primary-antibodies/cathepsin-l-v-k-h-antibody-epr8011-ab133641'>ab133641</a>) at 1/1000 dilution

Lane 5:

Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/1000 dilution

Lane 6:

Anti-His tag at 1/10000 dilution

All lanes:

Human Cathepsin V recombinant protein fraction

Secondary

Lanes 1 - 3:

Peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/1000 dilution

Lanes 4 - 5:

Rabbit Anti-Mouse secondary ab at 1/2000 dilution

Lane 6:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 38 kDa

false