SerpinB9 Overexpression Lysate

Name: SerpinB9 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-15851

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-15851

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for SerpinB9 Overexpression Lysate

SerpinB9 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC200645

Accession#: NM_004155

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

42.2 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for SerpinB9 Overexpression Lysate

Western Blot: SerpinB9 Overexpression Lysate [NBL1-15851]

Western Blot: SerpinB9 Overexpression Lysate (Adult Normal) [NBL1-15851] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for SerpinB9.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: SerpinB9

SerpinB9 is a serine proteinase inhibitor of the ovalbumin like B clade of serpins. The mouse orthologue to PI9 is SPI-6. SerpinB9 was first discovered in human bone marrow in a search for serpins similar to SerpinB6. SerpinB9 was identified in lymphocytes, especially natural killer cells and cytotoxic T lymphocytes, cells which also produce and store the apoptosis-inducing enzyme Granzyme B. PI9 was shown to be a potent inhibitor of Granzyme B, working at picamolar efficiencies. PI9 was also shown to inhibit caspase 1, the interleukin 1 converting enzyme, thus blocking IL1 223; production, and decreasing inflammatory processes. Most leukemia cells and many tumor cells produce PI9, leading to speculation that PI9 may help protect tumor cells from destruction by NK and CTL cells. PI9 is also found in placenta, lung, kidney, mast cells, and many tissues and cell lines. In kidney transplant, PI9 is sharply elevated, and in the liver PI9 has been shown to be upregulated by a downstream estrogen promoter. SerpinB9 inhibits Granzyme B, and is cleaved by Granzyme M, as a down regulatory step. SerpinB9 also inhibits the bacterial proteinase subtilisin A, and this may be a protective agent against infections. Neutrophil elastase is also a target of SerpinB9. Like SerpinB6 and SerpinB8, SerpinB9 lacks a signal sequence, and is found mainly in the cytoplasm and nucleus, although it can be detected outside of cells and in serum. The original SerpinB9 sequence described was 379 amino acids in length, with predicted mass of 42.4 kDa and pI of 5.51.

Alternate Names

CAP-3, CAP3PI-9, Cytoplasmic antiproteinase 3, Peptidase inhibitor 9, PI9protease inhibitor 9 (ovalbumin type), serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 9, serpin B9, serpin peptidase inhibitor, clade B (ovalbumin), member 9, serpin peptidase inhibitor, clade B, member 9

Gene Symbol

SERPINB9

Additional SerpinB9 Products

Product Documents for SerpinB9 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for SerpinB9 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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