CXCR5 Recombinant Rabbit Monoclonal Antibody [JB11-40]
cat.: ET7107-74
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC, IF-Cell
Clonality: Monoclonal
Clone number: JB11-40
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 42 kDa
Isotype: IgG
Immunogen: Synthetic peptide within C-terminal human CXCR5.
Positive control: Daudi cell lysate, EL4 cell lysate, Neuro-2a cell lysate, C6 cell lysate, mouse spleen tissue lysate, rat spleen tissue lysate, Daudi, Neuro-2a, rat kidney tissue, human tonsil tissue, mouse heart tissue
Subcellular location: Cell membrane.
Recommended Dilutions:
  WB
  IHC-P
  FC
  IF-Cell
1:2,000-1:5,000
1:50-1:200
1:50-1:100
1:250
Uniprot #: SwissProt: P32302 Human | Q04683 Mouse | P34997 Rat
Alternative names: BLR 1 BLR1 Burkitt lymphoma receptor 1 Burkitt lymphoma receptor 1 GTP binding protein Burkitt lymphoma receptor 1, GTP binding protein (chemokine (C-X-C motif) receptor 5) C X C chemokine receptor type 5 C-X-C chemokine receptor type 5 CD 185 CD185 CD185 antigen Chemokine (C-X-C motif) receptor 5 Chemokine C X C motif receptor 5 Chemokine CXC motif receptor 5 Chemokine receptor 5 CXC chemokine receptor type 5 CXC R5 CXC-R5 CXCR 5 CXCR-5 Cxcr5 CXCR5_HUMAN MDR 15 MDR-15 MDR15 MGC117347 Monocyte derived receptor 15 Monocyte-derived receptor 15
Images
ET7107-74_1.jpg Fig1: Western blot analysis of CXCR5 on different lysates with Rabbit anti-CXCR5 antibody (ET7107-74) at 1/2,000 dilution.

Lane 1: Daudi cell lysate
Lane 2: EL4 cell lysate
Lane 3: Neuro-2a cell lysate
Lane 4: C6 cell lysate
Lane 5: Mouse spleen tissue lysate
Lane 6: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

Exposure time: 1 minute 34 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-74) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
ET7107-74_2.jpg Fig2: Immunocytochemistry analysis of Daudi cells labeling CXCR5 with Rabbit anti-CXCR5 antibody (ET7107-74) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CXCR5 antibody (ET7107-74) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET7107-74_3.jpg Fig3: Immunocytochemistry analysis of Neuro-2a cells labeling CXCR5 with Rabbit anti-CXCR5 antibody (ET7107-74) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CXCR5 antibody (ET7107-74) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET7107-74_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-CXCR5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-74, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET7107-74_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CXCR5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-74, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET7107-74_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-CXCR5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-74, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET7107-74_7.jpg Fig7: Flow cytometric analysis of CXCR5 was done on Daudi cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-74, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.