Recombinant
RabMAb

Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641)

Overview

  • Product name
    Anti-Cathepsin L/V/K/H antibody [EPR8011]
  • Description
    Rabbit monoclonal [EPR8011] to Cathepsin L/V/K/H
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cathepsin L/V/K/H aa 100-200.
    Database link: P07711

  • Positive control
    • WB: A549, HCT-116, NIH/3T3, PC-12 and HepG2 cell lysates. IHC-P: Human kidney tissue. ICC/IF: HepG2 cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab133641 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.
WB 1/10000. Predicted molecular weight: 38 kDa.

For unpurified use at 1/1000 - 1/10000.

IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified use at 1/50 - 1/100.

Flow Cyt Use at an assay dependent concentration.

Images

  • Antigen: CATL_HUMAN, CATV_HUMAN, CATS_HUMAN, CATK_HUMAN, CATH_HUMAN

    Antigen concentration: 1000ng/ml

    Primary antibody concentration range: 0~1000ng/ml

    Secondary antibody: Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)

    Secondary antibody concentration: 1/2500

  • ab133641 staining Cathepsin L in the human cell line HepG2 (human hepatocellular carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • 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

    Lane 1 : Human Cathepsin L recombinant protein fraction
    Lane 2 : Human Cathepsin L recombinant protein fraction
    Lane 3 : 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 IgG 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

  • 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

    Lane 1 : Human Cathepsin V recombinant protein fraction
    Lane 2 : Human Cathepsin V recombinant protein fraction
    Lane 3 : Human Cathepsin V recombinant protein fraction
    Lane 4 : Human Cathepsin V recombinant protein fraction
    Lane 5 : Human Cathepsin V recombinant protein fraction
    Lane 6 : 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

  • Direct ELISA antigen dose-response curve using purified ab133641.Antigen concentration of 1000ng/mL.Alkaline Phosphatase conjugated AffiniPure goat anti-rabbit IgG(H+L)(1/2500) was used as the secondary antibody.

  • Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/20000 dilution (purified) + HepG2 cell lysate at 20 µg

    Secondary
    Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 38 kDa
    Observed band size : 38 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/10000 dilution (purified)

    Lane 1 : HCT-116 cell lysate
    Lane 2 : A549 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 38 kDa
    Observed band size : 38 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/20000 dilution (purified)

    Lane 1 : NIH/3T3 cell lysate
    Lane 2 : PC-12 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 38 kDa
    Observed band size : 38 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/1000 dilution (unpurified)

    Lane 1 : A549 cell lysate
    Lane 2 : HCT-116 cell lysate
    Lane 3 : NIH/3T3 cell lysate
    Lane 4 : PC-12 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 38 kDa
  • Anti-Cathepsin L/V/K/H antibody [EPR8011] (ab133641) at 1/1000 dilution (unpurified) + HepG2 cell lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 38 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Cathepsin L with purified ab133641 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Cathepsin L with unpurified ab133641 at a 1/50 dilution.

  • Immunocytochemistry/Immunofluorescence analysis of HT-29 cells labelling Cathepsin L with purified ab133641 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Unpurified ab133641 staining Cathepsin L in Human HepaRG cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% milk for 30 minutes at room temperature. Samples were incubated with primary antibody (1/1000) for 30 minutes. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Mizunoe Y  et al. Involvement of lysosomal dysfunction in autophagosome accumulation and early pathologies in adipose tissue of obese mice. Autophagy 13:642-653 (2017). Read more (PubMed: 28121218) »
  • Jacomin AC  et al. A functional endosomal pathway is necessary for lysosome biogenesis in Drosophila. BMC Cell Biol 17:36 (2016). ICC/IF ; Drosophila melanogaster . Read more (PubMed: 27852225) »

See all 5 Publications for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
10 µg
Sample
Trypanosoma brucei
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (HepaRG)
Specification
HepaRG
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jun 25 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 1µg/mL · Temperature: RT°C
Sample
Human Cell (HepG2)
Specification
HepG2
Permeabilization
Yes - 0.1% Triton X-100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 25 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Sample
Human Cell (HepaRG)
Specification
HepaRG
Permeabilization
Yes - 0.1% Triton x-100
Fixative
Formaldehyde
Username

Mrs. Alina Macovei

Verified customer

Submitted May 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HepaRG)
Loading amount
20 µg
Specification
HepaRG
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

Mrs. Alina Macovei

Verified customer

Submitted May 15 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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