CRABP1 Overexpression Lysate

Western Blot: CRABP1 Overexpression Lysate (Adult Normal) [NBP2-07877] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CRABP1.

• Reactivity: Hu
• Applications: WB

CRABP1 Overexpression Lysate Summary

• Description:

  CRABP1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC223810
  
  Accession#: NM_004378
  
  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.
• Gene: CRABP1

Applications/Dilutions

• Dilutions:
  • Western Blot
• 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.
• Theoretical MW: 15.4 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.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

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.

Alternate Names for CRABP1 Overexpression Lysate

• cellular retinoic acid binding protein 1
• CRABP
• CRABPI
• CRABP-Icellular retinoic acid-binding protein 1
• RBP5Cellular retinoic acid-binding protein I

Background

Retinoic acid (RA), a metabolite of vitamin A, plays an important role in growth and differentiation by modulation of expression of certain genes. RA functions through its interaction with the nuclear protein, retinoic acid receptor (RAR) and the 9-cis RA with retinoid X receptor (RXR). The Cellular Retinoic Acid Binding Proteins (CRABP) bind RA with a high affinity and belong to the family of fatty acid binding proteins. Two forms of CRABP have been characterized, CRABP 1 and CRABP 2 which share ~75% sequence conservation. The main role of CRABP 1 may be to protect retinoids from non-specific dehydrogenases and direct them to specific enzymes. The precise role of CRABP 2 is still poorly understood but it seems to play a role in the interaction of ligands (RA, 9-cis RA) with nuclear receptors (RAR/RXR).

Limitations

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.

Bioinformatics

• Gene Symbol: CRABP1