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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Rat Cystatin B Antibody

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.

Detection of Human Cystatin B by Simple Western^TM.

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.

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.

Applications for Human/Rat Cystatin B Antibody

Application

Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human bladder, colon, and heart

Immunoprecipitation

25 µg/mL
Sample: 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.

Simple Western

10 µg/mL
Sample: A549 human lung carcinoma cell line

Western Blot

1 µg/mL
Sample: 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

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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

Abrahamson, M. (1994) Methods Enzymol. 244:685.
Pol, E. and I. Bjork (1999) Biochemistry 38:10519.
Pennacchio, L.A. et al. (1996) Science 271:1731.
Zhang, R. et al. (2003) Glia 42:194.
Zerovnik, E. et al. (2002) Biochim. Biophys. Acta 1594:1.

Alternate Names

CSTB, Stefin B

Entrez Gene IDs

1476 (Human); 13014 (Mouse)

Gene Symbol

CSTB

UniProt

P04080

Additional Cystatin B Products

Product Documents for Human/Rat Cystatin B Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human/Rat Cystatin B Antibody

For research use only

Citations
Reviews

Human/Rat Cystatin B Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1408

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications