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CXCR4 Antibody

Catalog # NB100-74396 | Novus Biologicals a Bio-Techne Brand
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NB100-74396
NB100-74396SS

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Product Summary for CXCR4 Antibody

Immunogen

A synthetic peptide made to a C-terminal region of the human CXCR4 protein (within residues 300-352). [Swiss-Prot P61073]

Reactivity Notes

Rat reactivity reported in scientific literature (PMID:33116799).

Localization

Cell membrane; Multi-pass membrane protein. Note: In unstimulated cells, diffuse pattern on plasma membrane. On agonist stimulation, colocalizes with ITCH at the plasma membrane where it becomes ubiquitinated.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for CXCR4 Antibody

Immunocytochemistry/Immunofluorescence: CXCR4 Antibody [NB100-74396] - Optimization of immunofluorescence staining of CXCR4 protein. MDA, PC3, KG1 cells were used for the optimization experiments. Cells were seeded onto cell-tak coated 48 mm coverslips in a 48-well plate. Cells were fixed with 2% formaldehyde for 20 min, washed with PBS. After fixation, cells were blocked with 2.0% BSA in PBS for 1 h at room temperature. All the cells were incubated with a primary antibody for anti-rabbit CXCR4 for 1 h. After washing with PBS, cells were put in respective secondary antibodies-anti-rabbit dylight 405 for 1 h. PSMA= Magenta, EpCAM= Yellow, sLex= Green, CXCR4= Red, and Merge shows all the colors. MDA=PSMA+, EpCAM+, sLex+, CXCR4+. PC3 = PSMA-, EpCAM+, sLex-, CXCR4+. KG1 = PSMA-, EpCAM-, sLex+, CXCR4-. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0085143) licensed under a CC-BY license.
Western Blot: CXCR4 Antibody [NB100-74396] - The A2AR and baicalin attenuated CXCR4 expression in the hypoxia-induced PAH mouse model. CXCR4 protein expression levels in lung tissue were examined by western blot. GAPDH served as an internal control (c, d; n = 3). Image collected and cropped by CiteAb from the following publication (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5543745/) licensed under a CC-BY license.
Immunohistochemistry: CXCR4 Antibody [NB100-74396] - Pep R54 in combination with nivolumab reduced the expression of CXCR4-CXCL12 and PD-L1 in PES43 tumors. Representative IHC pictures (magnification 400x) for CXCR4, CXCR7 (red staining), CXCL12, PD-1 and PD-L1 expression (brown staining) in PES43 collected tumors from mice treated with Pep R54, nivolumab or combined treatment. Image collected and cropped by CiteAb from the following publication (https://jeccr.biomedcentral.com/articles/10.1186/s13046-019-1420-8) licensed under a CC-BY license.

Applications for CXCR4 Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50

Immunohistochemistry

1:200-1:1000

Immunohistochemistry-Paraffin

1:200-1:1000

Western Blot

1:5000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 3 using NB100-74396 in the following applications:

Published Applications

Read 29 publications using NB100-74396 in the following applications:

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Whole antisera

Preservative

0.05% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

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: CXCR4

CXCR4 (C-X-C chemokine receptor type 4) is a member of GPCR1 family and is expressed as a multipass membrane protein in several tissues wherein it acts as a receptor for the C-X-C chemokine CXCL12/SDF-1 that transduces a signal by increasing intracellular calcium ion levels and enhancing MAPK1/MAPK3 activation. CXCR4 acts as a receptor for extracellular ubiquitin; leading to enhanced intracellular calcium ions and reduced cellular cAMP levels. CXCR4 is involved in hematopoiesis, cardiac ventricular septum formation and plays an essential role in vascularization of the gastrointestinal tract via regulating vascular branching and/or remodeling processes in endothelial cells. CXCR4 is involved in cerebellar development and mediates hippocampal-neuron survival in CNS. It also acts as a coreceptor (CD4 being the primary receptor) for HIV-1 X4 isolates and as a primary receptor for some HIV-2 isolates, and promotes Env-mediated fusion of the virus.

Long Name

C-X-C Motif Chemokine Receptor 4

Alternate Names

CD184, D2S201E, FB22, Fusin, HM89, HSY3RR, LAP-3, LAP3, LCR1, LESTR, NPY3R, NPYRL, NPYY3R, WHIMS

Entrez Gene IDs

7852 (Human); 12767 (Mouse)

Gene Symbol

CXCR4

UniProt

Product Documents for CXCR4 Antibody

Certificate of Analysis

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

Product Specific Notices for CXCR4 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 CXCR4 Antibody (NB100-74396):

[[URL:https://www.novusbio.com/products/cxcr4-antibody_nb100-74396]][[Caption… Antibody]]
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 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 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%.


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.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution 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 biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

FAQs for CXCR4 Antibody

Showing  1 - 1 of 1 FAQ Showing All
  • Q: Which is your best CXCR4 for immunohistochemistry in paraffin tissues? I would like to detect it in paraffin embedded tissues from human breast cancer samples.

    A: CXCR4 antibody (NB100-74396) is our best selling product among all the CXCR4 antibodies and it has been validated for IHC-P in human cervical carcinoma tissue sections. NB100-74396 has been cited in at least 7 research publications. Additionally, here is a list of all of our CXCR4 antibodies that has been verified in IHC-P in human samples.

Showing  1 - 1 of 1 FAQ Showing All
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