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  1. Link

    cystatin-c-antibody-epr4413-ab109508.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cystatin C antibody [EPR4413] (ab109508)

  • Datasheet
Reviews (2) Submit a question References (22)

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Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
  • Anti-Cystatin C antibody [EPR4413] (ab109508)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4413] to Cystatin C
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-Cystatin C antibody [EPR4413]
    See all Cystatin C primary antibodies
  • Description

    Rabbit monoclonal [EPR4413] to Cystatin C
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human testis, Caco2, U20S, HeLa, HepG2, and U87-MG lysates, rat heart tissue lysate and mouse thymus tissue lysate. IHC-P: Human brain and kidney tissue, mouse kidney tissue and rat kidney tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4413
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Inhibitors
    • Stem Cells
    • Neural Stem Cells
    • Extracellular
    • Cardiovascular
    • Atherosclerosis
    • Thrombosis
    • Other
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-Cystatin C antibody [EPR4413] - BSA and Azide free (ab217569)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Mouse Cystatin C protein (ab63286)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109508 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)
1/10000 - 1/50000. Detects a band of approximately 13 kDa (predicted molecular weight: 15 kDa).
IHC-P
1/250 - 1/8000.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 13 kDa (predicted molecular weight: 15 kDa).
IHC-P
1/250 - 1/8000.
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IP.

Target

  • Function

    As an inhibitor of cysteine proteinases, this protein is thought to serve an important physiological role as a local regulator of this enzyme activity.
  • Tissue specificity

    Expressed in submandibular and sublingual saliva but not in parotid saliva (at protein level). Expressed in various body fluids, such as the cerebrospinal fluid and plasma. Expressed in highest levels in the epididymis, vas deferens, brain, thymus, and ovary and the lowest in the submandibular gland.
  • Involvement in disease

    Amyloidosis 6
    Macular degeneration, age-related, 11
  • Sequence similarities

    Belongs to the cystatin family.
  • Post-translational
    modifications

    The Thr-25 variant is O-glycosylated with a core 1 or possibly core 8 glycan. The signal peptide of the O-glycosylated Thr-25 variant is cleaved between Ala-20 and Val-21.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P01034 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1471 Human
    • Entrez Gene: 13010 Mouse
    • Entrez Gene: 25307 Rat
    • Omim: 604312 Human
    • SwissProt: P01034 Human
    • SwissProt: P21460 Mouse
    • SwissProt: P14841 Rat
    • Unigene: 304682 Human
    • Unigene: 4263 Mouse
    • Unigene: 106351 Rat
    see all
  • Alternative names

    • AD 8 antibody
    • AD8 antibody
    • Amyloid angiopathy and cerebral hemorrhage antibody
    • ARMD11 antibody
    • bA218C14.4 (cystatin C) antibody
    • bA218C14.4 antibody
    • Cst 3 antibody
    • Cst3 antibody
    • CST3 protein antibody
    • Cystatin 3 antibody
    • Cystatin-3 antibody
    • Cystatin-C antibody
    • Cystatin3 antibody
    • CystatinC antibody
    • CYTC_HUMAN antibody
    • Epididymis secretory protein Li 2 antibody
    • Gamma trace antibody
    • Gamma-trace antibody
    • HCCAA antibody
    • HEL S 2 antibody
    • MGC117328 antibody
    • Neuroendocrine basic polypeptide antibody
    • Post gamma globulin antibody
    • Post-gamma-globulin antibody
    see all

Images

  • Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: CST3 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Caco2 whole cell lysate (20 µg)
    Lane 4: U20S whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109508 observed at 15 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab109508 was shown to specifically react with CST3 when CST3 knockout samples were used. Wild-type and CST3 knockout samples were subjected to SDS-PAGE. Ab109508 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 10000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)

    Immunohistochemistry analysis of paraffin-embedded human kidney tissue sections labelling Cystatin C with ab109508 at 1/8000 dilution. The section was incubated with ab109508 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Positive staining on human kidney tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Anti-Cystatin C antibody [EPR4413] (ab109508) at 1/10000 dilution + Mouse thymus tissue lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 15 kDa
    Observed band size: 13 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking buffer and concentration : 5% NFDM/TBST
    Diluting buffer and concentration : 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)

    Immunohistochemistry analysis of paraffin-embedded rat kidney tissue sections labelling Cystatin C with ab109508 at 1/8000 dilution. The section was incubated with ab109508 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Positive staining on rat kidney tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)

    Immunohistochemistry analysis of paraffin-embedded mouse kidney tissue sections labelling Cystatin C with ab109508 at 1/8000 dilution. The section was incubated with ab109508 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Positive staining on mouse kidney tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Anti-Cystatin C antibody [EPR4413] (ab109508) at 1/10000 dilution + Rat heart tissue lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 15 kDa
    Observed band size: 13 kDa why is the actual band size different from the predicted?


    Exposure time: 80 seconds


    Blocking buffer and concentration : 5% NFDM/TBST
    Diluting buffer and concentration : 5% NFDM/TBST

  • Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Western blot - Anti-Cystatin C antibody [EPR4413] (ab109508)
    All lanes : Anti-Cystatin C antibody [EPR4413] (ab109508) at 1/10000 dilution

    Lane 1 : Human testis lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : U87-MG cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 15 kDa
    Observed band size: 13 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin C antibody [EPR4413] (ab109508)

    Immunohistochemical analysis of paraffin-embedded Human brain tissue using ab109508 at adilution of 1/250.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-Cystatin C antibody [EPR4413] (ab109508)
    Anti-Cystatin C antibody [EPR4413] (ab109508)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (22)

Publishing research using ab109508? Please let us know so that we can cite the reference in this datasheet.

ab109508 has been referenced in 22 publications.

  • Indacochea A  et al. Cold-inducible RNA binding protein promotes breast cancer cell malignancy by regulating Cystatin C levels. RNA 27:190-201 (2021). PubMed: 33172965
  • Wang M  et al. Deciphering the autophagy regulatory network via single-cell transcriptome analysis reveals a requirement for autophagy homeostasis in spermatogenesis. Theranostics 11:5010-5027 (2021). PubMed: 33754041
  • Shen Y  et al. Pressure overload promotes cystatin C secretion of cardiomyocytes to regulate the MAPK signaling pathway and mediate cardiac hypertrophy. Ann Transl Med 8:1514 (2020). PubMed: 33313259
  • Grigore F  et al. BRAF inhibition in melanoma is associated with the dysregulation of histone methylation and histone methyltransferases. Neoplasia 22:376-389 (2020). PubMed: 32629178
  • Kogot-Levin A  et al. Proximal Tubule mTORC1 Is a Central Player in the Pathophysiology of Diabetic Nephropathy and Its Correction by SGLT2 Inhibitors. Cell Rep 32:107954 (2020). PubMed: 32726619
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-Cystatin C antibody [EPR4413]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - other (concentrated secreted media of HeLa cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
1e+006 cells
Specification
concentrated secreted media of HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Dr. Daniele Stalder

Verified customer

Submitted Oct 06 2021

Immunohistochemistry (Frozen sections) abreview for Anti-Cystatin C antibody [EPR4413]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.1 M PBS with 3% Triton X
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Read More

DR. Ruma Raha-Chowdhury

Verified customer

Submitted Aug 20 2015

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