Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µg 298 185 ₸

Order now and get it on Thursday March 04, 2021

Anti-Cathepsin L + V antibody [33/2] (ab6314)

Key features and details

Mouse monoclonal [33/2] to Cathepsin L + V
Suitable for: WB, IHC-P, Flow Cyt
Reacts with: Mouse, Rat, Human
Isotype: IgG1

Product name

Anti-Cathepsin L + V antibody [33/2]
See all Cathepsin L + V primary antibodies
Description

Mouse monoclonal [33/2] to Cathepsin L + V
Host species

Mouse
Specificity

Reacts specifically with the native and denatured forms of human cathepsin L (25kD) and procathepsin L (42 kD). Does not react with human cathepsin types B1, D1, H and S1, procathepsins B and D1 and recombinant procathepsin H, rat cathepsin B1, and mouse procathepsins B1 and D1.

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Full length native protein (purified) corresponding to Human Cathepsin L + V.
Epitope

Recognizes an epitope within amino acid residues GYGFEST (265-271 in procathepsin L and 169-175 in the mature cathepsin L molecule).
Positive control
- This antibody gave a positive signal in human and mouse lung tissue lysates and in human, mouse and rat kidney tissue lysates. In Flow Cytometry, this antibody gave a positive result in methanol fixed/Tween permeabilised HeLa cells. IHC-P: FFPE human colon adenocarcinoma tissue sections.
General notes

This antibody clone is manufactured by Abcam.

If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine

Some batches contain 6.97% L-Arginine as a stabilizing agent. For lot-specific buffer information, please contact our Scientific Support team.
Concentration information loading...
Purity

Immunogen affinity purified
Clonality

Monoclonal
Clone number

33/2
Isotype

IgG1
Research areas

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

Lane 1 : Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/20 dilution
Lane 2 : 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 & 3 : Peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/1000 dilution
Lane 2 : Rabbit Anti-Mouse secondary ab at 1/2000 dilution

Predicted band size: 38 kDa

Blocking buffer and concentration:5% NFDM/TBST

Diluting buffer and concentration:5% NFDM/TBST

Observed MW:30

Exposure time:3 minutes
Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314)

Lane 1 : Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/20 dilution
Lane 2 : Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/100 dilution
Lane 3 : Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/500 dilution
Lane 4 : Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/100 dilution
Lane 5 : 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

Blocking buffer and concentration:5% NFDM/TBST

Diluting buffer and concentration:5% NFDM/TBST

Observed MW:30

Exposure time:3 minutes
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
Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314)

All lanes : Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/1000 dilution (3% milk block)

Lane 1 : Lung (Human) Tissue Lysate
Lane 2 : Lung (Mouse) Tissue Lysate
Lane 3 : Human kidney tissue lysate - total protein (ab30203)
Lane 4 : Kidney (Mouse) Tissue Lysate
Lane 5 : Kidney (Rat) Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 38 kDa
Observed band size: 25 kDa
why is the actual band size different from the predicted?
Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.

Exposure time: 20 minutes

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.
Western blot - Anti-Cathepsin L + V antibody [33/2] (ab6314) Image courtesy of Ratnakar Singh by Abreview.

Anti-Cathepsin L + V antibody [33/2] (ab6314) at 1/750 dilution + whole cell lysate prepared from U87MG human glioblastoma at 50 µg

Secondary
Goat anti-mouse HRP at 1/2000 dilution

Predicted band size: 38 kDa

See Abreview
Flow Cytometry - Anti-Cathepsin L + V antibody [33/2] (ab6314)

Overlay histogram showing HeLa cells stained with ab6314 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6314, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

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