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CCR5 Antibody, Novus Biologicals

Bio-Techne includes Novus Biologicals | Catalog # NBP2-31374

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NBP2-31374
NBP2-31374SS

Key Product Details

Species Reactivity

Human, Mouse, Monkey

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

1.0 mg/ml

Product Summary for CCR5 Antibody

Immunogen

Partial recombinant protein made to an N-terminal portion of the human CCR5 protein (between amino acids 1-350)

Predicted Species

Primate (99%). Backed by our 100% Guarantee.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 21305529). Rhesus macaque reactivity reported in a verified customer review.

Localization

Multi-pass membrane protein with extra-cellular and cytoplasmic domains

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for CCR5 Antibody

Western Blot: CCR5 Antibody [NBP2-31374] - Image of anti-CCR5 antibody. Whole cell protein from Daudi (1), K562 (2) and THP-1 (3) cell lines were separated by SDS-PAGE and protein transfered to PVDF. The membrane was probed with anti-CCR5 antibody at 2 ug/mL and detected with an anti-rabbit-HRP secondary antibody and chemiluminescence. CCR5 is shown at 40 kDa.
Immunohistochemistry-Paraffin: CCR5 Antibody [NBP2-31374] - Detection of CCR5 and virus on brain tissue. CCR5 expression was analyzed using Immunohistochemistry on the layer III of frontal cortex from macaques that were uninfected (A) and untreated controls, and Meth-only treated (B). Pictures were at 16x magnification. Sites of interest (rectangles) were further magnified. Image collected and cropped by CiteAb from the following publication (http://bmcimmunol.biomedcentral.com/articles/10.1186/s12865-016-0145-0), licensed under a CC-BY license.
Western Blot: CCR5 Antibody [NBP2-31374] - Detection of CCR5 partial recombinant protein by using CCR5 antibody at a concentration of 0.5 ug/mL.

Applications for CCR5 Antibody

Application
Recommended Usage

Immunohistochemistry

5 ug/ml

Immunohistochemistry-Paraffin

5 ug/ml

Simple Western

1:50

Western Blot

2 ug/ml
Application Notes
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4.5 using NBP2-31374 in the following applications:

Published Applications

Read 3 publications using NBP2-31374 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

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: CCR5

The CC chemokine receptor 5 (CCR5) is a member of the CC-chemokine receptor family, and has the characteristic structure of a 7 transmembrane G protein-coupled receptor (GPCR). CCR5 regulates trafficking and effector functions of memory/effector Th1 cells, macrophages, NK cells, and immature dendritic cells. CCR5 and its ligands play an important role in viral pathogenesis. CCR5 represents the co-receptor for macrophage (M) and dual (T cell and M)-tropic immunodeficiency viruses. Together with the CD4 binding receptor, CCR5 plays a critical role in HIV entry into the target cells. Moreover, the CCR5 ligands macrophage inflammatory protein (MIP)-1 alpha, MIP-1 beta and RANTES act as endogenous inhibitors of HIV infection, making both CCR5 and its chemokine ligands attractive therapeutic targets for HIV infection. Recent studies have also highlighted the role of CCR5 in a variety of other human diseases, ranging from infectious and inflammatory diseases to cancer.

Alternate Names

CCR5, CD195

Entrez Gene IDs

1234 (Human)

Gene Symbol

CCR5

UniProt

Additional CCR5 Products

Product Documents for CCR5 Antibody

Certificate of Analysis

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

Product Specific Notices for CCR5 Antibody

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.

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Protocols

View specific protocols for CCR5 Antibody (NBP2-31374):

CCR5 Antibody:
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.
CCR5 Antibody:
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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