Shop Antibodies
Best Seller

Antibody Sequencing & Cloning Service

$3,150.00

Item Cat No.: BCABSE

Antibody: Monoclonal Antibody

Application: Sequencing and cloning

Reactivity: Human, rabbit, rodent

BiCell Scientific’s antibody sequencing & cloning service provides full-length sequencing of custom antibody from hybridoma cell line and reassembly into mammalian antibody expression vector.

Determining an antibody’s sequence is crucial for various downstream applications, including antibody engineering, affinity maturation, functional optimization, and database documentation.

BiCell Scientific’s antibody sequencing & cloning service provides full-length sequencing of custom antibody from hybridoma cell line using 5′-RACE PCR technique. The fully sequenced antibody will then be cloned into a mammalian expression vector for recombinant antibody production.

Steps Timeline

(weeks)

Deliverables

Internal testing milestone

QC standard
Phase I: Total RNA extraction from the hybridoma cell pellet
(Total RNA will be extracted and purified from hybridoma cell pellet)
1 week / COA

 

 

 

 

Phase II: Reverse transcription
(Total RNA will be transcribed into cDNA using either an Oligo(dT) or a CH/CL antisense primer.

 

 

1-2 weeks / COA
Phase III: 5′ RACE amplification of heavy and light chains
(For 5’RACE PCR, heavy and light chains will be amplified with our proprietary adaptor (sense primer) and a gene specific (CH/CL, reverse primer). PCR products will include the sequence of the signal peptide, variable domains and constant domains up to the reverse primer)

 

 

2-3 weeks / COA
Phase IV: Cloning into sequencing vector
(PCR products will be gel purified and cloned into a sequencing vector for sequencing)
1-2 weeks Detailed report including the sequence alignments of the heavy and light chains sent to customer

 

Sequencing

Customers will receive the following deliverables at the end of each project:

  • Fully sequenced Fv domains of heavy chain and light chain of customer’s antibody
  • Cloning Fv domains of heavy chain and light chain into pFUSE series of mammalian Fc expression vectors

For Research Use Only. Not for use in clinical diagnostics or therapeutics.

  • Total RNA extraction from the hybridoma cell pellet – 1-2 weeks
  • Reverse transcription  – 1-2 weeks
  • 5′ RACE amplification of heavy and light chains – 2-3 weeks
  • Cloning into sequencing vector – 1-2 weeks
  • Sequencing 5 clones – 1-2 weeks
  • Cloning Fv domain into pFUSE series of Fc domain – 1-2 weeks

Reviews

There are no reviews yet.

Be the first to review “Antibody Sequencing & Cloning Service”

"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

Contact us for questions or custom requests!