Anti-Cathepsin L + V抗体[33/2] (ab6314)

概述

  • 产品名称Anti-Cathepsin L + V抗体[33/2]
  • 描述
    小鼠单克隆抗体[33/2] to Cathepsin L + V
  • 特异性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.
  • 经测试应用适用于: WB, ELISA, IHC-Fr, ICC/IF, IHC-P, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

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

  • 表位Recognizes an epitope within amino acid residues GYGFEST (265-271 in procathepsin L and 169-175 in the mature cathepsin L molecule).
  • 阳性对照
    • 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.
  • 常规说明

    Alternative versions available:

    Anti-Cathepsin L antibody (HRP) [33/2] (ab197278)

性能

应用

Our Abpromise guarantee covers the use of ab6314 in the following tested applications.

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

应用 Ab评论 说明
WB 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 38 kDa). Abcam recommends using 3% milk as the blocking agent.
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/50.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

  • 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 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

  • 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

  • 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

  • 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
    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.
  • 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

    Image courtesy of Ratnakar Singh by Abreview.

    See Abreview

  • ab6314 at a 1/150 dilution staining Cathepsin L in CHO cells by Immunocytochemistry/ Immunofluorescence.
  • 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.

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

This product has been referenced in:
  • Novakovic P  et al. Relationships of CD163 and CD169 positive cell numbers in the endometrium and fetal placenta with type 2 PRRSV RNA concentration in fetal thymus. Vet Res 47:76 (2016). IHC . Read more (PubMed: 27494990) »
  • Brings S  et al. Diabetes-induced alterations in tissue collagen and carboxymethyllysine in rat kidneys: Association with increased collagen-degrading proteinases and amelioration by Cu(II)-selective chelation. Biochim Biophys Acta 1852:1610-8 (2015). WB ; Rat . Read more (PubMed: 25900786) »

See all 10 Publications for this product

Product Wall

Application Western blot
Sample Mouse Cell lysate - whole cell (macrophage cells)
Gel Running Conditions Reduced Denaturing
Loading amount 80 µg
Specification macrophage cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 30 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (kidney, lung)
Specification kidney, lung
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Dec 22 2014

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (mammary gland)
Specification mammary gland
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization No
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 3.2% · Temperature: 22°C
Username

Dr. Irina Teplova

Verified customer

提交于 Jul 19 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Glioblastoma)
Loading amount 50 µg
Specification Glioblastoma
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 30°C
Username

Dr. Ratnakar Singh

Verified customer

提交于 Jun 10 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (MDA MB 231 cells)
Loading amount 10 µg
Specification MDA MB 231 cells
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Apr 09 2009

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