Overview

  • Product nameAnti-Stefin B antibody
    See all Stefin B primary antibodies
  • Description
    Rabbit polyclonal to Stefin B
  • SpecificityDetects endogenous levels of total Stefin B protein.
  • Tested applicationsSuitable for: WB, ELISA, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide derived from human Stefin B.

  • Positive control
    • WB: A549, A431, COLO205 and HCT116 whole cell lysates. IHC-P: Human normal placenta.

Properties

Applications

Our Abpromise guarantee covers the use of ab53725 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
WB 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).
ELISA 1/20000.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration. PMID 18667530

Target

  • FunctionThis is an intracellular thiol proteinase inhibitor. Tightly binding reversible inhibitor of cathepsins L, H and B.
  • Involvement in diseaseDefects in CSTB are the cause of progressive myoclonic epilepsy type 1 (EPM1) [MIM:254800]. EPM1 is an autosomal recessive disorder characterized by severe, stimulus-sensitive myoclonus and tonic-clonic seizures. The onset, occurring between 6 and 13 years of age, is characterized by convulsions. Myoclonus begins 1 to 5 years later. The twitchings occur predominantly in the proximal muscles of the extremities and are bilaterally symmetrical, although asynchronous. At first small, they become late in the clinical course so violent that the victim is thrown to the floor. Mental deterioration and eventually dementia develop.
  • Sequence similaritiesBelongs to the cystatin family.
  • Cellular localizationCytoplasm. Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • CHROW21 antibody
    • CPI B antibody
    • CPI-B antibody
    • CST 6 antibody
    • CST6 antibody
    • CSTB antibody
    • Cystatin B (stefin B) antibody
    • Cystatin B antibody
    • Cystatin-B antibody
    • CYTB antibody
    • CYTB_HUMAN antibody
    • EPM1 antibody
    • EPM1A antibody
    • Liver thiol proteinase inhibitor antibody
    • PME antibody
    • Stefin-B antibody
    • STF B antibody
    • STFB antibody
    • ULD antibody
    see all

Anti-Stefin B antibody images

  • ICC/IF image of ab53725 stained MCF-7 cells. The cells were 100% methanol fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53725, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Ab53725 staining Human normal placenta. Staining is localized to cytoplasmic and nuclear compartments.
    Left panel: with primary antibody at 1ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • All lanes : Anti-Stefin B antibody (ab53725) at 1 µg/ml

    Lane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : COLO 205 (Human colon adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 23 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

References for Anti-Stefin B antibody (ab53725)

This product has been referenced in:
  • Wang X  et al. Cystatin B is a progression marker of human epithelial ovarian tumors mediated by the TGF-ß signaling pathway. Int J Oncol 44:1099-106 (2014). WB, IHC-P ; Human . Read more (PubMed: 24452274) »
  • Polajnar M  et al. Gain in toxic function of stefin B EPM1 mutants aggregates: correlation between cell death, aggregate number/size and oxidative stress. Biochim Biophys Acta 1843:2089-99 (2014). Human . Read more (PubMed: 24909779) »

See all 4 Publications for this product

Product Wall

Thank you for your reply and those precisions. We have an alternative anti-CSTB, ab92449, directed against the C-terminal region of the human protein (between amino acids 73 and 98). www.abcam.com/ab92449 The homology between the region...

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Thank you for contacting us and for reporting the problem you experienced with the antibody ab53725. I am sorry that the product you received did not perform as expected. Since at Abcam we are concern about the quality of the products w...

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