Custom antibody work-up in the core facility

The Johns Hopkins Oncology Tissue Services Immunostaining Core Facility performs custom antibody work-up. Related fees include a $100 set-up fee per antibody (covers: Initial consultation, experiment setup and administrative costs) plus $43 per slide for the optimization run(s) conducted during antibody work-up and eventual staining of the actual slides in question. The investigator supplies the antibodies. Each case is unique and fees may change depending on the scope of the request. Quotes can be provided.

Immunostaining is notoriously prone to generating false negative and false positive results. Unless there are well validated antibodies available for the target of interest we STRONGLY recommend that the investigators source robust positive and negative controls to be used during the initial antibody work-ups.

Examples of cell line controls for antibody validation:

Gene knock-outs and knock-ins

Cells treated with agents well known to significantly change expression of the target protein

siRNA-treated cells to significantly lower expression

Cells transfected with expression vectors, such as plasmids or lentivirus, etc.

Examples of tissue controls for antibody validation:

Tissues that are well documented to endogenously express no/low versus high amounts of the target

Mouse knock-out or knock-in lines

Diseased versus normal tissues with known differences in expression of the target

Without such rigorous controls it can be very difficult to interpret any staining eventually generated on your actual tissues of interest.

Furthermore, if there are no known well validated antibodies for your target, we advise trying multiple different antibodies during the work-up phase in order to increase the probability of finding one that performs well..

A typical antibody workflow may look like this:

1. Consult with investigator

2. Obtain candidate antibody and source positive and negative control cells or tissues

3. Perform initial staining run on controls

4. Use information from initial run to conduct additional optimization/validation run(s)

5. Once antibody has been validated and optimal conditions are determined for staining, run pilot staining runs on the investigator's tissue of interest

There are no guaranteed results from using this service.^* Fees will be accessed on work done and not outcome. Please provide primary antibody spec sheets with requisition. For best results please have your tissue processed and sectioned in our TMA/Histology Core facility.

^* Each case is unique and fees may change depending on the scope of the request. For the development of a new antibody, we charge based on the number of slides stained to develop the antibody, rather than provide a single price, as antibody development can vary extensively. Quotes can be provided.

Non-JHU affiliates should send an inquiry to Helen Fedor.

Prices listed are for JHU Cancer Center members and are subject to change. Prices for non-cancer center members can be found here

Consultation
Service Fee ¹	$100.00
Consultation (per 1/4 hour)²	$50.00


IHC/IF/CISH
Single antibody immunostain (per slide) - Researcher Provided³	$43.00
Single antibody immunostain (per slide) - OTS Core Provided⁴	$53.00
CISH (per slide)⁵	$130.00


Controls
Control slide - single tissue	$10.00
Control slide - Tissue Microarray	Inquire
Custom cell line array	Inquire


Misc.
Flat slide holder	$8.50
Microscope consult (per 1/4 hr.)	$25.00
Rush Fee
IHC can be rushed on any established antibodies, which will incur a 50% increase in the billing of the project.	.

1) Set up fee includes an initial consultative meeting, administrative and experimental setup costs. There is no initial consultation cost for antibodies that have been worked up in the core facility (antibodies listed below).
2) Consultation - assistance in finding antibodies, appropriate controls, etc.
3) Includes cost of all non-antibody reagents, technician time and consumables.
4) Includes cost of the antibody and all non-antibody reagents, technician time and consumables.
5) Includes cost of reagents, technician time and consumables. Investigator must supply the probe.