Anti-CXCR4 antibody [UMB2] - BSA and Azide free

15 product images

• 

  Fischer, T. et al PLoS One. 2008;3(12):e4069. doi: 10.1371/journal.pone.0004069. Epub 2008 Dec 31 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Characterization of UMB-2 (Anti-CXCR4 antibody [UMB2] ab124824) by immunofluorescent staining of transfected cells. HEK-293 cells expressing CCR7 or CXCR4 were either not exposed or exposed to 100 ng/ml MIP-3 or 100 ng/ml SDF-1 for 30 min, subsequently fixed and immunofluorescently stained with 1 µg/ml anti-CCR7 {1188} or anti-CXCR4 {UMB-2} at a dilution of 1:100. Note that UMB-2 detected prominent immunofluorescence at the level of the plasma membrane only in CXCR4- but not in CCR7-expressing cells, and that SDF-1 exposure induced a rapid translocation of CXCR4 receptor immunostaining from the plasma membrane into the cytosol. Representative results from one of three independent experiments are shown. Scale bar, 20 µm.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

• 

  Flow Cytometry (Intracellular) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Intracellular Flow Cytometry analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CXCR4 with purified Anti-CXCR4 antibody [UMB2] ab124824 at 1/260 dilution (10 μg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) - Black. Unlabeled control - Blue. Untreated cells - GreenThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197203)

• 

  Costa, M.J. et al PLoS One. 2018 Mar 19;13(3):e0194688. doi: 10.1371/journal.pone.0194688. eCollection 2018 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Immunohistochemical detection of CXCR4 expression in human tissue specimens of normal appearance
  

  CXCR4 was detected in the indicated PFA-fixed, paraffin-embedded human tissues using Anti-CXCR4 antibody [UMB2] ab124824 at 5 μ/ml overnight at 4°C.

  A, kidney. B, adrenal gland. C, cerebellum. D, bone marrow, brown staining: CXCR4, green staining: CD45. E, Spleen. F, testis. G, lung. H, colon.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Immunofluorescence staining of Jurkat cells with purified Anti-CXCR4 antibody [UMB2] ab124824 at a working dilution of 1 in 250, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and Alexa Fluor^® 594 goat anti-mouse at a dilution of 1/500 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) . The secondary antibody was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Alexa Fluor^® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified Anti-CXCR4 antibody [UMB2] ab124824 was used at a dilution of 1/200 followed by an Alexa Fluor^® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and anti-rabbit secondary antibody (ab15007) were used.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Clone UMB2 (ab197203) has been successfully conjugated by Abcam. This image was generated using Anti-CXCR4 antibody [UMB2] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-CXCR4 antibody [UMB2] ab208128 for protocol details.

  Alexa Fluor® 488 Anti-CXCR4 antibody [UMB2] ab208128 staining CXCR4 in Jurkat cells. The cells were fixed with 4% formaldehyde (10 min) and then incubated in 1%BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated overnight at +4°C with Alexa Fluor® 488 Anti-CXCR4 antibody [UMB2] ab208128 at 1/100 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  This product also gave a positive signal under the same testing conditions in Jurkat cells fixed with 80% methanol (5 min).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Clone UMB2 (ab197203) has been successfully conjugated by Abcam. This image was generated using Anti-CXCR4 antibody [UMB2] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-CXCR4 antibody [UMB2] ab208129 for protocol details.

  Alexa Fluor® 647 Anti-CXCR4 antibody [UMB2] ab208129 staining CXCR4 in Jurkat cells. The cells were fixed with 80% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 647 Anti-CXCR4 antibody [UMB2] ab208129 at 1/50 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  This product also gave a positive signal under the same testing conditions in Jurkat cells fixed with 4% formaldehyde (10 min).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Immunohistochemical staining of paraffin embedded human bladder cancer with purified Anti-CXCR4 antibody [UMB2] ab124824 at a working dilution of 1/500. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Unpurified Anti-CXCR4 antibody [UMB2] ab124824, at 1/50 dilution, staining CXCR4 in paraffin-embedded Human cervical carcinoma tissue by immunohistochemistry.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Anti-CXCR4 antibody [UMB2] ab124824 stained Jurkat cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-CXCR4 antibody [UMB2] ab124824 at 5ug/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Unpurified Anti-CXCR4 antibody [UMB2] ab124824, at 1/50 dilution, staining CXCR4 in paraffin-embedded Human ovarian adenocarcinoma tissue by immunohistochemistry.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Unpurified Anti-CXCR4 antibody [UMB2] ab124824, at 1/50 dilution, staining CXCR4 in paraffin-embedded Human tonsil tissue by immunohistochemistry.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CXCR4 with purified ab197203 at 1/1000 dilution (1.067 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA Buffer, PH9 (Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684). Hematoxylin was used to counter stain. Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used.
  

  Membranous with weak cytoplasmic staining on human tonsil.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human B-cell non-hodgkin lymphoma tissue sections labeling CXCR4 with purified ab197203 at 1/1000 dilution (1.067 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA Buffer, PH9 (Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684). Hematoxylin was used to counter stain. Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used.
  

  Cytoplasmic and membranous staining on tumor cells of human B-cell non-Hodgkin lymphoma.

• 

  Flow Cytometry (Intracellular) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).
  Flow cytometry overlay histogram showing left Jurkat positive cells and right negative HepG2 stained with Anti-CXCR4 antibody [UMB2] ab124824 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-CXCR4 antibody [UMB2] ab124824) (1x 106 in 100μl at 1.0μg/ml (1/1900)) for 30min at 22°C.
  
  The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
  
  Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
  
  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
  
  This antibody gave a positive signal in Jurkat Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• 

  Flow Cytometry (Intracellular) - Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CXCR4 antibody [UMB2] ab124824).
  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney) cells transfected with a mix of human CXCR1, CXCR2, CXCR3 and CXCR5 expression vector containing a myc-his tag (Left) / HEK-293T transfected with a human CXCR4 expression vector containing a myc-his tag (Right) labelling CXCR4 with Anti-CXCR4 antibody [UMB2] ab124824 at 1/2000 dilution (0.1μg)/ Left and Right. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/3000 dilution was used as the secondary antibody.
  
  No cross-reactivity with CXCR1/CXCR2/CXCR3/CXCR5.