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Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2b Lambda Clone # 2D8

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for CCR2 Antibody (2D8) - BSA Free

Immunogen

A synthetic peptide made to an N-terminal portion of the human CCR2 protein (between residues 1-100)

Predicted Species

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

Localization

Cell membrane

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2b Lambda

Scientific Data Images for CCR2 Antibody (2D8) - BSA Free

Western Blot: CCR2 Antibody (2D8) [NBP2-35333] - Analysis of CCR2 [2D8] in Molt4 lysate
Immunohistochemistry-Paraffin: CCR2 Antibody (2D8) [NBP2-35333] - Analysis of FFPE tissue section of human esophageal squamous cell carcinoma (SCC) using CCR2 antibody (clone 2D8) at 5 ug/ml concentration. The cancer cells depicted strong cytoplasmic-membranous immunostaining, whereas the cells of tumor stroma were largely negative for CCR2 immunopositivity.
Immunohistochemistry-Paraffin: CCR2 Antibody (2D8) [NBP2-35333] - Analysis of FFPE tissue section of human breast using CCR2 antibody (clone 2D8) at 5 ug/ml concentration. The glandular cells in the lobules showed a specific cytoplasmic-membranous immunostaining, whereas, the inter-lobular connective tissue did not develop any staining for CCR2.

Applications for CCR2 Antibody (2D8) - BSA Free

Application
Recommended Usage

Immunohistochemistry

5 ug/ml

Immunohistochemistry-Paraffin

5 ug/ml

Western Blot

2 ug/ml
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

0.05% 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: CCR2

CCR2, or C C chemokine receptor type 2, is a receptor for several monocyte chemo attractant proteins (MCP1, MCP3, MCP4) which specifically mediate monocyte chemotaxis. CCR2 transduces such signals by increasing the intracellular level of calcium ions. For example, MCP1 is involved in monocyte infiltration in inflammatory diseases such as rheumatoid arthritis as well as in the inflammatory response against tumors. CCR2 is also an alternative coreceptor with CD4 for HIV1 infection. CCR2 has two isoforms and has been reported to be expressed in a wide variety of tissues, including blood, brain, heart, kidney, liver, lung, ovary, pancreas, spinal cord, spleen and thymus.

Alternate Names

CC-CKR-2, CCR2, CCR2B, CD192, CKR2, CKR2A, CMKBR2, FLJ78302, MCP-1-R

Entrez Gene IDs

729230 (Human)

Gene Symbol

CCR2

UniProt

Additional CCR2 Products

Product Documents for CCR2 Antibody (2D8) - 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 CCR2 Antibody (2D8) - 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 CCR2 Antibody (2D8) - BSA Free (NBP2-35333):

CCR2 Antibody (2D8):
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.
CCR2 Antibody (2D8):
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-CCR2 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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