Anti-CD4 antibody [EPR6855]

22 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Anti-CD4 antibody [EPR6855] (ab133616)
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with ab133616 at a dilution of 1:500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
  This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

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  Baratella et al PLoS Negl Trop Dis. 2017 Jan; 11(1): e0005285. Published online 2017 Jan 17. doi: 10.1371/journal.pntd.0005285

  Immunocytochemistry/ Immunofluorescence - Anti-CD4 antibody [EPR6855] (ab133616)

  HBZ is preferentially expressed in CD4+ T cells of HAM/TSP patient PH1624

  Confocal microscopy analysis of PBMC from HAM/TSP patient PH1624. (A) co-staining with the 4D4-F3 anti-HBZ mAb followed by Alexa Fluor 546-conjugated goat anti-mouse IgG1 antibody (red) and with the anti-CD4 mAb followed by Alexa Fluor 488-conjugated goat-anti-rabbit IgG antibody (green); upper panels, extended field; lower panels, enlarged field focused on the single cell depicted in the square of the left upper panel and positive for both CD4 and HBZ.

  CD4 was detected using ab133616 at 1/100 dilution.

  From Figure 6A of Baratella et al.

  Baratelle et al PLoS Negl Trop Dis. 2017 Jan; 11(1): e0005285. Published online 2017 Jan 17. doi: 10.1371/journal.pntd.0005285

  Reproduced under Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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  Flow Cytometry (Intracellular) - Anti-CD4 antibody [EPR6855] (ab133616)

  Human peripheral blood lymphocytes stained with unpurifiedab133616 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (unpurified ab133616, 1/100 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with purified ab133616 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

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  Western blot - Anti-CD4 antibody [EPR6855] (ab133616)

  All lanes: Western blot - Anti-CD4 antibody [EPR6855] (ab133616) at 1/1000 dilution

  Lane 1: THP-1 cell lysate at 10 µg

  Lane 2: Human fetal thymus lysate at 10 µg

  Lane 3: Human tonsil lysate at 10 µg

  Lane 4: Human lymph node lysate at 10 µg

  Secondary

  All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution

  Predicted band size: 51 kDa

  Observed band size: 51 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunocytochemistry analysis of THP-1 (Human monocytic leukemia monocyte) labeling CD4 with purified ab133616 at 1/100 dilution. Cells were fixed with 100% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.30 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
  Negative control: PBS instead of the primary antibody.

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  Western blot - Anti-CD4 antibody [EPR6855] (ab133616)

  Different batches of ab133616 were tested on THP-1 (Human monocytic leukemia monocyte) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 51 kDa.

  All lanes: Western blot - Anti-CD4 antibody [EPR6855] (ab133616)

  Predicted band size: 51 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymoma tissue labelling CD4 with purified ab133616 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling CD4 with ab133616 at a dilution of 1/100.
  

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

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Paraffin-embedded human spleen tissue stained for CD4 using ab133616 at 1/500 dilution in immunohistochemical analysis.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with unpurified ab133616 at a dilution of 1/100.
  

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD4 with unpurified ab133616.
  
  

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD4 with unpurified ab133616.
  
  

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymoma tissue labelling CD4 with unpurified ab133616.
  

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Negative control: no staining on human cerebrum.
  

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum showing no staining CD4 with purified ab133616 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Counterstained with hematoxylin.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Negative control: no staining on human pancreas.
  

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreas showing no staining CD4 with purified ab133616 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Counterstained with hematoxylin.

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  Western blot - Anti-CD4 antibody [EPR6855] (ab133616)

  Blocking and dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-CD4 antibody [EPR6855] (ab133616) at 1/5000 dilution

  Lane 1: Human fetal thymus tissue lysate at 20 µg

  Lane 2: Human tonsil tissue lysate at 20 µg

  Lane 3: THP-1 cell lysate at 20 µg

  Lane 4: HuT-78 cell lysate at 20 µg

  Secondary

  All lanes: HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

  Predicted band size: 51 kDa

  Observed band size: 51 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Tissue Microarrays stained for "Anti-CD4 antibody [EPR6855]" using "ab133616"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab133616 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-CD4 antibody [EPR6855] - BSA and Azide free ab181724).

  Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4 with Anti-CD4 antibody [EPR6855] - BSA and Azide free ab181724 at a concentration of 0.69 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. Anti-CD4 antibody [EPR6855] - BSA and Azide free ab181724 Anti-CD4 antibody [EPR6855] was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4with ab133616 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with OptiView DAB IHC Detection Kit followed by OptiView Amplification kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab133616 anti-CD4 antibody [EPR6855] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4with ab133616 at a concentration of 1.43 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. ab133616 anti-CD4 antibody [EPR6855] was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR6855] (ab133616)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-CD4 antibody [EPR6855] - BSA and Azide free ab181724).

  Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4 with Anti-CD4 antibody [EPR6855] - BSA and Azide free ab181724 at a concentration of 1.37 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). Anti-CD4 antibody [EPR6855] - BSA and Azide free ab181724 Anti-CD4 antibody [EPR6855] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.