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Demonstration of Labeling with Non-aggregating High-resolution Secondary Antibody Enzyme Conjugate

Demonstration of Labeling with Non-aggregating High-resolution Secondary Antibody Enzyme Conjugate

FFPE sections of mouse kidney were labeled with anti-Sglt2 antibody followed by horseradish peroxidase conjugated secondary antibody. (Image courtesy of Dr. Kim, UCSD)

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Non-aggregating High-resolution Secondary Antibody Enzyme Conjugate

$375.00

Antibody: Secondary Polyclonal Antibody

Concentration: 0.50 mg/ml

Application: IHC on Cryosection and FFPE

Reactivity: Rabbit, Rat, Mouse

200 µl size

Non-aggregating High-resolution Secondary Antibody Enzyme Conjugate allows colorimetric labeling of primary polyclonal or monoclonal antibodies.

Enzyme conjugated secondary antibodies allow location studies of target proteins at cellular levels with colorimetric techniques.

The most common approach for enzyme conjugation to secondary antibody is via maleimide-succinimidyl ester. However, enzyme conjugation alters the solubility of antibody molecules, rendering aggregation of secondary antibodies at subcellular levels, which create punctate signals on micrographs.

BiCell Scientific Inc has developed a proprietary conjugation approach that is able to maintain the solubility of enzyme conjugated antibodies, thus minimizing the aggregation of secondary antibody molecules and improve the spatial resolution of colorimetric signals.

Host/Isotype: Goat/IgG

Class: Polyclonal

Immunogen: Target species IgG (H+L)

Conjugation: Enzyme

Purification: Affinity chromatography

Storage buffer: PBS, pH 7.2, 0.1% sodium azide

Storage condition: –20°C

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

  • Deparaffinize FFPE sections with Xylene.
  • Rehydrate sections with decreasing concentrations of ethyl alcohol (100% to 70%).
  • Wash slides 2 times with dH2O.
  • Dip slides into universal antigen retrieval buffer.
  • Microwave sections at boiling temperature for 10min.
  • Wash slides 2 times with dH2O.
  • Dip slides into FFPE autofluorescence quenching buffer for 20min.
  • Wash slides with 70% ethyl alcohol
  • Wash slides with 1xPBS
  • Drain Wash solution against a towel and put slides into the wet chamber.
  • Dilute primary antibody at 1:50 in Block solution and add to each slide (300-500 µl is sufficient to cover the section on each slide).
  • Incubate the slides in the wet chamber at room temperature for 2hr.
  • Take slides out of the wet chamber, drain off antibody, and wash three times with the Wash solution.
  • Dilute secondary antibodies at 1: 200 in Block solution and add to each slide and put slides back into the wet chamber.
  • Incubate the slides in the wet chamber at room temperature for 1hr.
  • Take slides out of the wet chamber, drain off antibody, and wash three times with the Wash solution.
  • Follow substrate reaction protocol to develop colorimetric signals.

Additional information

Target species

Rabbit, Rat, Mouse
Enzyme conjugates

Horseradish Peroxidase, Alkaline Phosphatase

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"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

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