CRTH-2/GPR44 Overexpression Lysate

Western Blot: GPR44 Overexpression Lysate (Adult Normal) [NBL1-11281] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GPR44.

• Reactivity: Hu
• Applications: WB

CRTH-2/GPR44 Overexpression Lysate Summary

• Description:

  CRTH-2/GPR44 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC223378
  
  Accession#: NM_004778
  
  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: PTGDR2

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: 43.1 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 CRTH-2/GPR44 Overexpression Lysate

• CD294 antigen
• CD294
• Chemoattractant receptor-homologous molecule expressed on Th2 cells
• CRTH2
• CRTH-2
• CRTH2chemoattractant receptor homologous molecule expressed on T helper type 2 cells
• DL1R
• DP2
• G protein-coupled receptor 44
• GPR44
• PGD2R
• PTGDR2
• putative G-protein coupled receptor 44

Background

GPR44, also called CRTH2, is a Prostanoid Receptor that binds prostaglandin D2. PGD2 signals through CRTH2 to induce Th2, eosinophil, and basophil chemotaxis in a G-alpha(i) dependent manner. CRTH2 may play a role in mast cell-mediated immune responses such as those involved in allergic reactions. This receptor is upregulated in activated T helper-2 (Th2) cells, peripheral blood basophils, and eosinophils, but not in Th1 lineage cells. It may be a useful marker for Th2 and Tc2 cells. CRTH2 has been reported to be expressed in the central nervous system, peripheral tissues, and in blood cells, particularly activated T helper-2 cells. Two transcripts have been isolated, a 3.5-kb transcript expressed primarily in the thalamus, frontal cortex, pons, and hippocampus, and a 3.4-kb transcript expressed in fetal liver, leukocytes, and thymus. ESTs have been isolated from B-cell/lung/testis, brain, kidney, liver/spleen, placenta, prostate, testis, and thymus libraries.

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.

Publications for CRTH-2/GPR44 Lysate (NBL1-11281)(1)

Publication using NBL1-11281

• Qian F, Nettleford SK, Zhou J et al. Activation of GPR44 decreases severity of myeloid leukemia via specific targeting of leukemia initiating stem cells Cell reports 2023-07-25 [PMID: 37459233] (WB)

FAQs for CRTH-2/GPR44 Lysate (NBL1-11281). (Showing 1 - 1 of 1 FAQ).

1. I have couple of questions regarding the GPR44 lysate (NBL1-11281). 1. What is the primary antibody used in western blot analysis? 2. Can I use this lysate as control for any other GPR44 antibody (like from other company)? 3. Does my primary antibody has to be specific anti-DDK antibody (since it has DDK tag)?
   • In regards to your inquiry, 1) We use an anti DDK tag antibody to test in Western blot. 2) You can use this as a control for another antibody from another company. One thing to keep in mind however, is if the protein of interest naturally expresses in 293 cells, bands may still be detected in the empty vector control with an antibody against the GPR44 protein instead of the tag. 3)The primary does not have to be specific to DDK tag as the protein of interest should be expressed there, but note the image on the website is from anti-DDK antibody.

Bioinformatics

• Gene Symbol: PTGDR2