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Complete IHC kit for FFPE section

$850.00$950.00

Item Cat No.: BCASKP

Application: IHC on FFPE

Reactivity: Tissue

BiCell Scientific’s complete IHC kit for FFPE section provides a full set of reagents including two internal control antibodies for immunolabeling of FFPE sections using fluorescent or chromogenic detection method.

1 Kit (100 slides)

Formaldehyde crosslinks are formed between protein molecules, in particular with the basic amino acid – lysine. Formalin-fixed paraffin-embedded (FFPE) sections (formalin: 10% formaldehyde) preserve tissue morphology, but require antigen retrieval processes to revive antigenicity in tissues, which is essential to IHC applications.

BiCell Scientific’s complete IHC kit for FFPE section provides a full set of reagents and protocols including two internal control antibodies, rabbit anti E-cadherin polyclonal antibody and rat anti ZO-1 polyclonal antibody, for immunolabeling of FFPE sections using fluorescent or chromogenic detection method.

The kit components (fluorescent option):

  • 50 ul of anti E-cadherin antibody (rabbit polyclonal)
  • 50 ul of anti ZO-1 antibody (rat polyclonal)
  • 50 ul of anti rabbit secondary antibody (Alexa Fluor 488)
  • 50 ul of anti rat secondary antibody (Alexa Fluor 594)

Store at –20oC

  • 500 ml of Universal antigen retrieval buffer (proprietary)
  • 100 ml of FFPE autofluorescence quenching buffer (proprietary)
  • 100 ml of Block buffer (bovine serum, Tween-20, NaCl and Tris buffered saline)
  • 500 ml of Wash buffer (Tween-20, NaCl, and Tris buffered saline)
  • 1 ml of Antifade-Fluorescence Mounting Medium

Store at 4oC


The kit components (enzyme-substrate option):

  • 50 ul of anti E-cadherin antibody (rabbit polyclonal)
  • 50 ul of anti ZO-1 antibody (rat polyclonal)
  • 50 ul of anti rabbit secondary antibody (Horseradish peroxidase)
  • 50 ul of anti rat secondary antibody (Alkaline phosphatase)

Store at –20oC

  • 500 ml of Universal antigen retrieval buffer (proprietary)
  • 100 ml of Block buffer (bovine serum, Tween-20, NaCl and Tris buffered saline)
  • 500 ml of Wash buffer (Tween-20, NaCl, and Tris buffered saline)
  • 50 ml of DAB substrate for Horseradish peroxidase
  • 50 ml of BCIP/NBT substrate for Alkaline phosphatase
  • 10 ml of Antifade-Chromogen Mounting Medium

Store at 4oC


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

Fluorescent detection:

  • Thaw kit content to room temperature.
  • 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.
  • Add 20 ul of Antifade-fluorescence mounting medium to the section on each slide. Place a cover glass onto the section and apply a small pressure to remove any trapped air bubble. Air-dry at room temperature for 1hr. Store glass slides at 4oC for less than 1 month.

 


Enzyme-substrate detection:

  • Thaw kit content to room temperature.
  • 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.
  • 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.
  • Apply 100 ul of substrate solution to tissue section until desired color is achieved (1-10 mins).
  • Rinse off substrate with PBS.
  • Dehydrate sections with increasing concentrations of ethyl alcohol (70% to 100%).
  • Dehydrate sections with Xylene. Air-dry in fume hood for 30min at room temperature.
  • Add 100 ul of Antifade-chromogen mounting medium to the section on each slide. Place a cover glass onto the section and apply a small pressure to remove any trapped air bubble. Air-dry at room temperature for 1hr. Store glass slides at ambient temperature.

Additional information

Detection

Fluorescence, Enzyme-Substrate

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Be the first to review “Complete IHC kit for FFPE section”

"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

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