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Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2b Kappa Clone # 3A7.1B8

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for Cyr61/CCN1 Antibody (3A7.1B8) - BSA Free

Immunogen

Partial recombinant protein from human Cyr61/CCN1 protein (between residues 100-300). [Swiss-Prot O00622]

Predicted Species

Primate (100%). Backed by our 100% Guarantee.

Reactivity Notes

Immunogen's sequence similarity with other species: Rat (81%), Mouse (79%), Camel (88%), Naked Mole Rat (87%), Porcine (80%), Bovine / Sheep / Horse (82%), Chicken (63%)

Localization

Secreted (Cytoplasmic/Extracellular)

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2b Kappa

Scientific Data Images for Cyr61/CCN1 Antibody (3A7.1B8) - BSA Free

Western Blot: Cyr61/CCN1 Antibody (3A7.1B8) [NBP2-36490] - WB analysis of (A) 12kDa Partial Recombinant Cyr61/CCN1 protein and (B) Human Liver lysate using Cyr61/CCN1 antibody (clone 3A7.1B8) at 4ug/ml concentration.
Immunohistochemistry-Paraffin: Cyr61/CCN1 Antibody (3A7.1B8) [NBP2-36490] - IHC analysis of formalin-fixed paraffin-embedded tissue section of human liver using Cyr61/CCN1 antibody (clone 3A7.1B8) at 5 ug/ml concentration. The hepatocytes generated a specific cytoplasmic staining with no signal in the cellular nuclei or the sinusoids.
Immunohistochemistry-Paraffin: Cyr61/CCN1 Antibody (3A7.1B8) [NBP2-36490] - IHC analysis of formalin-fixed paraffin-embedded tissue section of human hepatocellular carcinoma using Cyr61/CCN1 antibody (clone 3A7.1B8) at 5 ug/ml concentration. The cancer cells generated a specific cytoplasmic staining with some staining in the inter-cellular spaces but no signal in the cellular nuclei, tumor stroma or the RBCs.

Applications for Cyr61/CCN1 Antibody (3A7.1B8) - BSA Free

Application
Recommended Usage

Immunohistochemistry

5 ug/ml

Immunohistochemistry-Paraffin

5 ug/ml

Western Blot

4 ug/ml
Application Notes
Protein CYR61 is a secreted protein and its expected immunolocalization would be cytoplasmic as well as the extracellular spaces.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Cyr61/CCN1

Cysteine-rich angiogenic inducer 61, also known as CYR61, is a growth factor-inducible, immediate-early gene that has multifaceted activities in various cancers. CYR61 is a secreted heparin-binding extracellular matrix-associated protein whose activities include the promotion of adhesion and chemotaxis, and the stimulation of fibroblast and endothelial cell growth. CYR61 induces angiogenesis and promotes tumor growth and is expressed in dermal fibroblasts during cutaneous wound healing.

Long Name

Cysteine-Rich, Angiogenic Inducer 61

Alternate Names

CCN1, GIG1, IGFBP-10

Entrez Gene IDs

3491 (Human); 16007 (Mouse); 83476 (Rat)

Gene Symbol

CCN1

UniProt

Additional Cyr61/CCN1 Products

Product Documents for Cyr61/CCN1 Antibody (3A7.1B8) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Cyr61/CCN1 Antibody (3A7.1B8) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Protocols

View specific protocols for Cyr61/CCN1 Antibody (3A7.1B8) - BSA Free (NBP2-36490):

Cyr61/CCN1 Antibody (3A7.1B8):
1. Deparaffinize the tissue sections by immersing the slides in Xylene with two changes for 10 min each. Sections should not get dried at any stage from this point.
2. Rehydrate the tissue sections by immersing the slides in decreasing grades of ethanol as follows:
a. Immerse in 100% ethanol with 2 changes for 5 minutes each
b. Immerse in 95% ethanol with 2 changes for 5 minutes each
c. Immerse in 90% ethanol for 5 minutes
d. Immerse in 70% ethanol for 5 minutes
e. Immerse in 50% ethanol for 5 minutes
f. Immerse in distilled water for 5 minutes
3. Antigen Retrieval (Microwave Method):
a. Immerse the slides in a microwave compatible tray containing 10 mM Sodium Citrate buffer (pH 6.0) with 0.05% Tween 20.
b. Boil the slides and maintain the sub-boiling temperature for 5 minutes in the microwave. Thereafter, take out the tray very carefully and cool it at room temperature (RT) for about 30 minutes.
c. Wash the slides 3 times, 3 minutes each by immersing them in TBST (Tris Buffered Saline having 0.05% Tween 20).
4. Quenching of Endogenous Peroxidase:
a. Incubate the slides in 3% hydrogen peroxide prepared in methanol for 15 minutes (at RT, in dark conditions).
b. Wash the slides in TBST 3 times, 3 minutes each.
5. Protein Blocking:
a. Incubate the sections with background sniper solution at RT for 15 minutes (Biocare Medicals, USA).
b. Wash the sections 3 times, 3 min each by immersing the slides in TBST.
6. Primary Antibody:
a. Dilute the primary antibody at 5ug/ml concentration using PBS as a diluent.
b. Incubate the sections with diluted primary antibody for 90 minutes at RT in a humidified chamber.
c. Thereafter, wash the slides 4 times, 5 minutes each with TBST.
7. Probe (Secondary Reagent):
a. Incubate with MACH 1 Mouse probe for 15 minutes at RT.
b. Incubate for 30 min at room temperature with HRP-Polymer (Biocare Medical, USA).
c. Wash the slides with TBST 4 times, 5 minutes each
8. Chromogen:
a. Mix 32ul of DAB Chromogen with 1 ml of DAB substrate buffer (Biocare Medical, USA).
b. Apply 200ul DAB mixture/section and incubate at RT in dark conditions (few seconds - 5 minutes).
c. As soon as an appropriate color develops, rinse the slides with deionized water (2-3 brief rinses).
9. Counter stain:
a. Counter stain with Hematoxylin for 30 seconds (Vector Labs, USA).
b. Wash in deionized water for 1-2 minutes to clear the extra stain.
c. Incubate the slides in bluing solution or Scott's water twice for 2 minutes each time.
10. Dehydrate the sections in increasing grades of alcohols:
a. 50% alcohol for 1 minute
b. 70% for 1 minute
c. 90% for 1 minute
d. 95% for 1 minute
e. 100% for 1 minute
f. Xylene with 2 changes for 2 minutes each
11. Mount with DPX mount and cover-slip glass (Fisher Scientific, USA), carefully not allowing any air bubbles to enter.

NOTE:- This protocol is provided as a reference tool only. Depending upon the type of tissues /tissue processing and reagents employed, the end user will need to optimize the final conditions for achieving an expected staining.
Cyr61/CCN1 Antibody (3A7.1B8):
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 25 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute anti-Cyr61/CCN1 (3A7.1B8) primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

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