CD74 Overexpression Lysate

Name: CD74 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-08968

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-08968

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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 CD74 Overexpression Lysate

CD74 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC212964

Accession#: NM_001025159

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

33.3 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 CD74 Overexpression Lysate

Western Blot: CD74 Overexpression Lysate [NBL1-08968]

Western Blot: CD74 Overexpression Lysate (Adult Normal) [NBL1-08968] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD74.

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: CD74

CD74, also known as the MHC class II associated invariant chain (Ii), is a type II transmembrane protein which binds to the peptide binding groove of newly synthesized MHC class II alpha/beta heterodimers and prevents their premature association with endogenous polypeptides. CD74 is produced in molar excess over MHC class II and some of this is expressed by an unknown pathway on the cell surface independent of, or in association with, MHC class II molecules. The half life of CD74 on the cell surface is only 3 to 4 min after which it is internalized. It has been proposed that cell surface CD74 binds MHC class II molecules which have lost their antigenic peptide leading to internalization and reloading with antigenic peptide. The transport route of class II alpha/beta/Ii complexes is regulated selectively by two forms of CD74 (p33 and p35) which are generated by the use of alternative translation initiation sites. CD74 is expressed primarily by antigen presenting cells such as B lymphocytes (from before the pre-B cell stage to before the plasma cell stage), macrophages and monocytes together, with many epithelial cells. CD74 may exist in different isoforms ranging in size from 33 to 41 kDa, depending on genetic splicing.

Alternate Names

CD74, CLIP, DHLAG, HLADG, Ia-gamma, INVG34

Gene Symbol

CD74

Additional CD74 Products

Product Documents for CD74 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for CD74 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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