DHFR Monoclonal antibody

• 

  WB analysis using 68274-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68274-1-Ig (DHFR antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

• 

  WB analysis of NIH/3T3 using 68274-1-Ig

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 68274-1-Ig (DHFR antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  FC experiment of MCF-7 using 68274-1-Ig

  1x10^6 MCF-7 cells were intracellularly stained with 0.4 ug DHFR Monoclonal antibody (68274-1-Ig, Clone:4C11E9) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1)(red), or 0.4 ug Mouse IgG1 isotype control Mouse McAb (66360-1-Ig, Clone: 1F8D3) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

DHFR Monoclonal Antibody for WB, FC (Intra), ELISA

Cat No. 68274-1-Ig

Clone No.4C11E9

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse

Applications

WB, FC (Intra), ELISA

Synonyms

4C11E9, DHFRP1, EC:1.5.1.3

---

Tested Applications

• Positive WB detected in: LNCaP cells, NIH/3T3 cells, HeLa cells, HEK-293 cells, Jurkat cells
• Positive FC (Intra) detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

68274-1-Ig targets DHFR in WB, FC (Intra), ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag7318

  Product name: Recombinant human DHFR protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 1-187 aa of BC000192

  Sequence: MVGSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

• Full Name: dihydrofolate reductase
• Calculated Molecular Weight: 21 kDa
• Observed Molecular Weight: 21 kDa
• GenBank Accession Number: BC000192
• Gene Symbol: DHFR
• Gene ID (NCBI): 1719
• RRID: AB_2935357
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: P00374
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

DHFR (dihydrofolate reductase) catalyzes the subsequent NADPH-dependent reduction of the H2folate produced by TS to regenerate the tetrahydrofolate (H4folate) necessary for continued dTMP synthesis and transfer of 1-carbon units (PMID:3461439). It belongs to the dihydrofolate reductase family and can exist as a homodimer (PMID:2248959). DHFR is widely expressed in fetal and adult human brain and whole blood, expression is higher in adult brain than fetal brain (PMID:21310276).