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Location: Home > Custom Services > Antibody Discovery Services > IHC-Positive Hybridoma Screening

IHC-Positive Hybridoma Screening

Date: 2018-08-27 Author: Leading Biology Click: 415

Introduction

Immunohistochemical (IHC) staining is widely used in the diagnosis of abnormal cells such as those found in cancerous tumors. Specific molecular markers are characteristic of particular cellular events such as proliferation or cell death (apoptosis). Immunohistochemistry is also widely used in basic research to understand the distribution and localization of biomarkers and differentially expressed proteins in different parts of a biological tissue. Immunohistochemistry (IHC) involves the process of selectively imaging antigens (proteins) in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues.


Based on molecular experimental professional team, Leading Biology established a complete set of services for IHC fidelity antibody preparation, after obtaining a large number of suspected positive hybridoma cells, we could verify the IHC availability antibodies for customers, and hand the purified hybridoma cell supernatant to customers for further validation test. One the one hand, it saves the customers' work and shorten the test cycle, on the other hand, it eliminates the false positive problem caused by the risk of sample transportation. At present, in addition to providing IHC validation services, we also face a vast number of scientific research and medical facilities, providing IHC experimental service. According to the customer's needs, we could complete the entire experimental service process from tissue embedding, slicing, staining, filming and analyzing.


Service Package

Service Items

Time

(months)

Start point

Peptide

Small Molecule

 

Phosphate/Methyl

/Acetyl-specific

Purified protein

Expression plasmid

Prokaryotic proteins synthesis de novo

Eukaryotic proteins synthesis de novo

Peptide design & synthesis

Peptide design

0.5

Free, Optional

 

 

 

 

Peptide synthesis

 

 

 

 

 

Conjugation

 

 

 

 

Proteins preparation

cDNA synthesis

1

 

 

 

 

 

Need further inquiry

Need further inquiry

Vector construction

 

 

 

 

 

Protein expression & purification

 

 

 

 

Immunization

Immune 5 mice

1.5

ELISA test

Cell Fusion & Screening

1.5

SubcloningELISA Screening)

0.5

Monoclonal (IHC Screening)

1

Ascites production

(Inject 3 mice)

1

Purification and detection

Affinity purification

0.5

Antigen specific affinity purification

 

 

 

 

 

 

 

Total lead time(months)

4-6

-4

4-6

5-7

6-8


Classic Case

Customer Provide

Protein antigen\plasmid\protein information + 20 antibody positive paraffin sections (3-5μm)


Process

1. Antigen preparation\Plasmid expression\Protein purification\Peptide design, synthesis and conjugation.

2. Immunization of 5-6 Balb/c mice.

3. ELISA/Western-blot test to monitor the antibody titer and specificity in mice serum, according to ELISA and WB test result, screening mice.

4. Cell fusion: fusing with the myeloma cells SP2/0.

5. Positive screening: Using ELISA test to screening the antibody positive hybridoma clone.

6. Subclone: Using limited dilution to produce subclone from positive clone. According to customers’ requirements, we also could screen out the monoclonal cell line which could produce Ig antibodies firmly (extra charge).

7. 3 positive cell clones were screened out by using IHC, and the purified supernatants of cell cultures were sent to customers for test.

8. Ascites preparation: Using mice celiac inoculation to produce the ascites of hybridoma products.

9. Antibody purification: Purify the secretory antibodies from 2 monoclonal hybridoma cell lines.


Products 

(each antigen, 5 mice)

1. 5mg polypeptide

2. 2 immunogen specific monoclonal hybridoma cell lines.

3. Each cell line provide 10ml (1mg/ml) purified antibody, ELISA  titer was above 1:50,000.WB test positive.

4. If customer requires, we could provide: prepared ascites (titer 1:500-1000, extra charge), hybridoma cell culture supernatant (for WB test).

5. Complete lab report, remaining antigens.


Preparation Period

4-5 months


Successful Result

Publication: Res Vet Sci.

IF: 2.191

Title: Changes in the expression of Heat Shock Proteins in ovaries from bovines with cystic ovarian disease induced by ACTH.

Name: anti-HSP27/HSP60/HSP70/HSP90 antibody

Species: Argentinean Holstein heifers

Type: Mouse monoclonal antibody


Related Services

Mouse Monoclonal Antibody Service

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Monkey Monoclonal Antibody Service

Canine Monoclonal Antibody Service

Hamster Hybridoma Production


Why Leading Biology

With over 5 years of experience in custom antibodies, we offer numerous benefits for developing your custom antibody:


• Our industry leading titer guarantees of 1:50,000 eliminate the risk of not obtaining antibodies against peptide antigens.

• Our economies of scale allow us to pass cost savings to you and help maximize your budget.

• 100% of services are transparent throughout.

• Antibodies from our facility have been cited in many published research papers.

• By outsourcing antibody production and characterization needs to us, you can spend more time focused on your research.

• Our ideal location in California ensures that animals benefit from outdoor facilities and mild year-round temperatures.

• Confidentiality is a top priority for all projects. Antibodies that we manufacture belong to you and will not be commercialized.


Contact Information

Please obtain a quote before ordering, and refer to the quote number when you place an order.
Orders are typically confirmed within 12 hours.
Have a Question? Email us info@leadingbiology.com
Order Products: Order Related Products
By Phone: 1-661-524(LBI)-0262 (USA)


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