Band 3 Overexpression Lysate

Name: Band 3 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-11143

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-11143

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Product Datasheet / COA / SDS

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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 Band 3 Overexpression Lysate

Band 3 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC218312

Accession#: NM_000342

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

101.6 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 Band 3 Overexpression Lysate

Western Blot: Band 3 Overexpression Lysate [NBP2-11143]

Western Blot: Band 3 Overexpression Lysate (Native) [NBP2-11143] - Band 3 Overexpression Lysate (Native) Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for SLC4A1/AE1.

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: Band 3

The protein encoded by the Band 3 gene is part of the anion exchanger (AE) family and is expressed in the erythrocyte plasma membrane, where it functions as a chloride/bicarbonate exchanger involved in carbon dioxide transport from tissues to lungs. The protein comprises two domains that are structurally and functionally distinct. The N-terminal 40kDa domain is located in the cytoplasm and acts as an attachment site for the red cell skeleton by binding ankyrin. The glycosylated C-terminal membrane-associated domain contains 12-14 membrane spanning segments and carries out the stilbene disulphonate-sensitive exchange transport of anions. The cytoplasmic tail at the extreme C-terminus of the membrane domain binds carbonic anhydrase II. The encoded protein associates with the red cell membrane protein glycophorin A and this association promotes the correct folding and translocation of the exchanger. This protein is predominantly dimeric but forms tetramers in the presence of ankyrin. Many mutations in this gene are known in man, and these mutations can lead to two types of disease: destabilization of red cell membrane leading to hereditary spherocytosis, and defective kidney acid secretion leading to distal renal tubular acidosis. Other mutations that do not give rise to disease result in novel blood group antigens, which form the Diego blood group system. Southeast Asian ovalocytosis (SAO, Melanesian ovalocytosis) results from the heterozygous presence of a deletion in the encoded protein and is common in areas where Plasmodium falciparum malaria is endemic. One null mutation in this gene is known, resulting in very severe anemia and nephrocalcinosis. (provided by RefSeq)

Alternate Names

AE 1, AE1MGC126619, Anion exchange protein 1, Anion exchanger 1, band 3 anion transport protein, BND3, CD233, CD233 antigen, DI, EMPB3, EPB3MGC126623, erythrocyte membrane protein band 3, erythroid anion exchange protein, FR, Froese blood group, MGC116750, MGC116753, RTA1A, Solute carrier family 4 member 1, solute carrier family 4, anion exchanger, member 1 (erythrocyte membraneprotein band 3, Diego blood group), solute carrier family 4, anion exchanger, number 1, SW, Swann blood group, Waldner blood group, WD, WD1, WR, Wright blood group

Gene Symbol

SLC4A1

Additional Band 3 Products

Product Documents for Band 3 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for Band 3 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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