Home / Cystatin B / Human/Rat Cystatin B Antibody

Human/Rat Cystatin B Antibody

Catalog #: AF1408
3 Citations Datasheet / COA / SDS
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AF1408
AF1408-SP
Detection of Human and Rat Cystatin B by Western Blot.
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Citations (3)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Rat Cystatin B Antibody Summary

Species Reactivity
Human, Rat
Specificity
Detects human and rat Cystatin B in Western blots. In Western blots, less than 2% cross-reactivity with recombinant human (rh) Cystatin A, recombinant mouse Cystatin B, rhCystatin C, rhCystatin D, rhCystatin E/M, rhCystatin S, rhCystatin SA, and rhCystatin SN is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Cystatin B
Met2-Phe98
Accession # P04080
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
Immersion fixed paraffin-embedded sections of human bladder, colon, and heart
Immunoprecipitation
25 µg/mL
Conditioned cell culture medium spiked with Recombinant Human Cystatin B (Catalog # 1408-PI), see our available Western blot detection antibodies
Knockout Validated
Cystatin B is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in Cystatin B knockout HEK293T cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human and Rat Cystatin B antibody by Western Blot. View Larger

Detection of Human and Rat Cystatin B by Western Blot. Western blot shows lysates of human colon tissue, PC-12 rat adrenal pheochromocytoma cell line, A549 human lung carcinoma cell line, and SK-BR-3 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Rat Cystatin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1408) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Cystatin B at approximately 11 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western Detection of Human Cystatin B antibody by Simple Western<SUP>TM</SUP>. View Larger

Detection of Human Cystatin B by Simple WesternTM. Simple Western lane view shows lysates of A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Cystatin B at approximately 18 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Rat Cystatin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1408) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Knockout Validated Western Blot Shows Human Cystatin B Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human Cystatin B Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and Cystatin B knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Rat Cystatin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1408) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Cystatin B at approximately 12 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Cystatin B

Cystatin B, also called stefin B or liver thiol proteinase inhibitor, is a member of family 1 of the cystatin superfamily (1). Like Cystatin A, it is an intracellular inhibitor regulating the activities of cysteine proteases of the papain family such as cathepsins B, H and L (2). Mutations in the Cystatin B gene is the cause of progressive myoclonus epilepsy (EPM1) (3). Because of its expression patterns, Cystatin B can be used as a marker for certain cancers, such as glioblastoma tumors (4). It readily forms amyloid fibrils in vitro (5). The human Cystatin B consists of 98 amino acid residues (3).

References
  1. Abrahamson, M. (1994) Methods Enzymol. 244:685.
  2. Pol, E. and I. Bjork (1999) Biochemistry 38:10519.
  3. Pennacchio, L.A. et al. (1996) Science 271:1731.
  4. Zhang, R. et al. (2003) Glia 42:194.
  5. Zerovnik, E. et al. (2002) Biochim. Biophys. Acta 1594:1.
Entrez Gene IDs
1476 (Human); 13014 (Mouse)
Alternate Names
CPI-B; CST6cystatin B (liver thiol proteinase inhibitor)10STFBcystatin-B; CSTB; cystatin B (stefin B); Cystatin B; EPM1; Liver thiol proteinase inhibitor; PME; Stefin B; stefin-B

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Citations for Human/Rat Cystatin B Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Notch signalling drives synovial fibroblast identity and arthritis pathology
    Authors: K Wei, I Korsunsky, JL Marshall, A Gao, GFM Watts, T Major, AP Croft, J Watts, PE Blazar, JK Lange, TS Thornhill, A Filer, K Raza, LT Donlin, CW Siebel, CD Buckley, S Raychaudhu, MB Brenner
    Nature, 2020-04-22;582(7811):259-264.
  2. Modulation of Receptor Protein Tyrosine Phosphatase Sigma Increases Chondroitin Sulfate Proteoglycan Degradation through Cathepsin B Secretion to Enhance Axon Outgrowth
    Authors: AP Tran, S Sundar, M Yu, BT Lang, J Silver
    J. Neurosci., 2018-05-14;38(23):5399-5414.
    Species: Rat
    Sample Types: Cell Culture Supernates, Whole Cells
    Applications: ICC, Western Blot
  3. Regulation of TGF-beta1-driven differentiation of human lung fibroblasts: emerging roles of cathepsin B and cystatin C.
    Authors: Kasabova M, Joulin-Giet A, Lecaille F, Gilmore B, Marchand-Adam S, Saidi A, Lalmanach G
    J Biol Chem, 2014-04-30;289(23):16239-51.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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