Skip to main content

Novus Biologicals products are now on bio-techne.com

Better as one

CCR2 Antibody - BSA Free

Catalog # NBP1-48338 | Novus Biologicals a Bio-Techne Brand

Key Product Details

Species Reactivity

Human, Mouse

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1 mg/ml

Product Summary for CCR2 Antibody - BSA Free

Immunogen

Synthetic peptide made to an N-terminal portion of the mouse CCR2 protein (within residues 20-100). [Swiss-Prot# P51683]

Localization

Cell membrane; Multi-pass membrane protein.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for CCR2 Antibody - BSA Free

Flow Cytometry: CCR2 Antibody [NBP1-48338] - An intracellular stain was performed on THP-1 cells with CCR2 Antibody NBP1-48338G (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 488.
Immunocytochemistry/Immunofluorescence: CCR2 Antibody [NBP1-48338] - Analysis of CCR2 in NIH/3T3 cells.
Immunohistochemistry: CCR2 Antibody [NBP1-48338] - Expression patterns of CCL2 and CCR2 in unwounded and wounded corneas of WT and MMP12 KO mice. Immunofluorescence of CCL2 chemokine receptor CCR2 in unwounded and chemically wounded (2-days after injury) WT and Mmp12-/- mouse corneas. Control images represent mouse corneas stained with secondary antibody only and without primary antibody. Nuclei were visualized by staining with DAPI (blue). Scale bars: 50 um. CCR2 staining was visualized in epithelial and stromal layers of unwounded and wounded WT and Mmp12-/- corneas. Image collected and cropped by CiteAb from the following publication (http://www.nature.com/articles/s41598-019-47831-z), licensed under a CC-BY license.

Applications for CCR2 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50

Immunohistochemistry

1:200

Immunohistochemistry-Frozen

reported in scientific literature (PMID 31399604)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 11 publications using NBP1-48338 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: CCR2

CCR2 is a major chemokine receptor expressed on monocytes and its activation through its ligands MCP1, MCP2, MCP3 and MCP4 (systematic names CCL2, CCL8, CCL7 and CCL13, respectively) stimulates the migration of monocytes across the vascular wall into tissues wherein they facilitate chronic inflammation. CCR2 is a member of GPCR1 family and is up-regulated by CREB3. It is a multi-pass membrane protein found in cell membrane, and besides monocytes, it is also found on other cell types like NK cells, macrophages, immature dendritic cells, gamma delta T cells, and activated T cells (including Th17 cells). There are two alternatively spliced forms of CCR2: CCR2A - the major isoform expressed by mononuclear cells and vascular smooth muscle cells; and CCR2B - expressed predominantly by monocytes and activated NK cells. CCR2 has both pro-inflammatory (mediated by APC and T cells) and anti-inflammatory (mediated by regulatory T cells) activities. Besides inflammation, it has been implicated several pathologies including HIV-1/AIDS, autoimmune disorders (psoriasis, rheumatoid arthritis, and multiple sclerosis), pulmonary diseases (asthma, COPD), vascular disease and cancer (prostate cancer).

Alternate Names

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

Entrez Gene IDs

12772 (Mouse)

Gene Symbol

CCR2

UniProt

Additional CCR2 Products

Product Documents for CCR2 Antibody - 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 - 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.

Loading...

Protocols

View specific protocols for CCR2 Antibody - BSA Free (NBP1-48338):

Protocol for Flow Cytometry Intracellular Staining
Sample Preparation.
1. Grow cells to 60-85% confluency. Flow cytometry requires between 2 x 105 and 1 x 106 cells for optimal performance.
2. If cells are adherent, harvest gently by washing once with staining buffer and then scraping. Avoid using trypsin as this can disrupt certain epitopes of interest. If enzymatic harvest is required, use Accutase, Collagenase, or TrypLE Express for a less damaging option.
3. Reserve 100 uL for counting, then transfer cell volume into a 50 mL conical tube and centrifuge for 8 minutes at 400 RCF.
a. Count cells using a hemocytometer and a 1:1 trypan blue exclusion stain to determine cell viability before starting the flow protocol. If cells appear blue, do not proceed.
4. Re-suspend cells to a concentration of 1 x 106 cells/mL in staining buffer (NBP2-26247).
5. Aliquot out 100 uL samples in accordance with your experimental samples.

Tip: When cell surface and intracellular staining are required in the same sample, it is advisable that the cell surface staining be performed first since the fixation and permeabilization steps might reduce the availability of surface antigens.

Intracellular Staining.
Tip: When performing intracellular staining, it is important to use appropriate fixation and permeabilization reagents based upon the target and its subcellular location. Generally, our Intracellular Flow Assay Kit (NBP2-29450) is a good place to start as it contains an optimized combination of reagents for intracellular staining as well as an inhibitor of intracellular protein transport (necessary if staining secreted proteins). Certain targets may require more gentle or transient permeabilization protocols such as the commonly employed methanol or saponin-based methods.
Protocol for Cytoplasmic Targets:
1. Fix the cells by adding 100 uL fixation solution (such as 4% PFA) to each sample for 10-15 minutes.
2. Permeabilize cells by adding 100 uL of a permeabilization buffer to every 1 x 106 cells present in the sample. Mix well and incubate at room temperature for 15 minutes.
a. For cytoplasmic targets, use a gentle permeabilization solution such as 1X PBS + 0.5% Saponin or 1X PBS + 0.5% Tween-20.
b. To maintain the permeabilized state throughout your experiment, use staining buffer + 0.1% of the permeabilization reagent (i.e. 0.1% Tween-20 or 0.1% Saponin).
3. Following the 15 minute incubation, add 2 mL of the staining buffer + 0.1% permeabilizer to each sample.
4. Centrifuge for 1 minute at 400 RCF.
5. Discard supernatant and re-suspend in 100 uL of staining buffer + 0.1% permeabilizer.
6. Add appropriate amount of each antibody (eg. 1 test or 1 ug per sample, as experimentally determined).
7. Mix well and incubate at room temperature for 30 minutes- 1 hour. Gently mix samples every 10-15 minutes.
8. Following the primary/conjugate incubation, add 1-2 mL/sample of staining buffer +0.1% permeabilizer and centrifuge for 1 minute at 400 RCF.
9. Wash twice by re-suspending cells in staining buffer (2 mL for tubes or 200 uL for wells) and centrifuging at 400 RCF for 5 minutes. Discard supernatant.
10. Add appropriate amount of secondary antibody (as experimentally determined) to each sample.
11. Incubate at room temperature in dark for 20 minutes.
12. Add 1-2 mL of staining buffer and centrifuge at 400 RCF for 1 minute and discard supernatant.
13. Wash twice by re-suspending cells in staining buffer (2 mL for tubes or 200 uL for wells) and centrifuging at 400 RCF for 5 minutes. Discard supernatant.
14. Resuspend in an appropriate volume of staining buffer (usually 500 uL per sample) and proceed with analysis on your flow cytometer.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


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

Loading...