Rabbit Monoclonal CD4 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, mIHC, Flow Cyt (Intra) and reacts with Human samples. Cited in 11 publications.
IgG
Rabbit
Constituents: PBS
Liquid
Monoclonal
IHC-P | ICC/IF | WB | mIHC | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes For unpurified use at 1/100 - 1/250. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info - | Notes For unpurified use at 1/100 - 1/250. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Please check the parent abID, Anti-CD4 antibody [EPR6855] ab133616, for a recommended dilution. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes For unpurified use at 1/1000 - 1/10000. |
Species Rat | Dilution info - | Notes For unpurified use at 1/1000 - 1/10000. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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Integral membrane glycoprotein that plays an essential role in the immune response and serves multiple functions in responses against both external and internal offenses. In T-cells, functions primarily as a coreceptor for MHC class II molecule:peptide complex. The antigens presented by class II peptides are derived from extracellular proteins while class I peptides are derived from cytosolic proteins. Interacts simultaneously with the T-cell receptor (TCR) and the MHC class II presented by antigen presenting cells (APCs). In turn, recruits the Src kinase LCK to the vicinity of the TCR-CD3 complex. LCK then initiates different intracellular signaling pathways by phosphorylating various substrates ultimately leading to lymphokine production, motility, adhesion and activation of T-helper cells. In other cells such as macrophages or NK cells, plays a role in differentiation/activation, cytokine expression and cell migration in a TCR/LCK-independent pathway. Participates in the development of T-helper cells in the thymus and triggers the differentiation of monocytes into functional mature macrophages.(Microbial infection) Primary receptor for human immunodeficiency virus-1 (HIV-1) (PubMed:2214026, PubMed:16331979, PubMed:9641677, PubMed:12089508). Down-regulated by HIV-1 Vpu (PubMed:17346169). Acts as a receptor for Human Herpes virus 7/HHV-7 (PubMed:7909607).
T-cell surface glycoprotein CD4, T-cell surface antigen T4/Leu-3, CD4
Rabbit Monoclonal CD4 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, mIHC, Flow Cyt (Intra) and reacts with Human samples. Cited in 11 publications.
T-cell surface glycoprotein CD4, T-cell surface antigen T4/Leu-3, CD4
IgG
Rabbit
Constituents: PBS
Liquid
Monoclonal
Yes
EPR6855
Affinity purification Protein A
Blue Ice
+4°C
Do Not Freeze
ab181724 is the carrier-free version of Anti-CD4 antibody [EPR6855] ab133616.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Human peripheral blood lymphocytes stained with unpurified Anti-CD4 antibody [EPR6855] ab133616 (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 Anti-CD4 antibody [EPR6855] ab133616, 1/100 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with purified Anti-CD4 antibody [EPR6855] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-CD4 antibody [EPR6855]. ab181724 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.
Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada Inc.
This data was developed using the same antibody clone in a different buffer formulation (Anti-CD4 antibody [EPR6855] ab133616).
Immunocytochemistry analysis of THP-1 (Human monocytic leukemia monocyte) labeling CD4 with purified Anti-CD4 antibody [EPR6855] 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.
This WB data was generated using the same anti-CD4 antibody clone [EPR6855] in a different buffer formulation (cat# Anti-CD4 antibody [EPR6855] ab133616).
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concetration: 5% NFDM/TBST
All lanes: Western blot - Anti-CD4 antibody [EPR6855] - BSA and Azide free (ab181724)
All lanes: THP-1 (human monocytic leukemia cell line) whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)
Predicted band size: 51 kDa
Exposure time: 3min
This data was developed using Anti-CD4 antibody [EPR6855] ab133616, the same antibody clone in a different buffer formulation. Different batches of Anti-CD4 antibody [EPR6855] 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] (Anti-CD4 antibody [EPR6855] ab133616)
Predicted band size: 51 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymoma tissue labelling CD4 with purified Anti-CD4 antibody [EPR6855] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Clone EPR6855 (ab181724) has been successfully conjugated by Abcam. This image was generated using Anti-CD4 antibody [EPR6855] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-CD4 antibody [EPR6855] ab196372 for protocol details.
Alexa Fluor® 488 Anti-CD4 antibody [EPR6855] ab196372 staining CD4 in Jurkat cells. The cells were fixed with 100% methanol (5 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 488 Anti-CD4 antibody [EPR6855] ab196372 at a working dilution of 1 in 50 (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling CD4 with Anti-CD4 antibody [EPR6855] ab133616 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Clone EPR6855 (ab181724) has been successfully conjugated by Abcam. This image was generated using Anti-CD4 antibody [EPR6855] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-CD4 antibody [EPR6855] ab196147 for protocol details.
Alexa Fluor® 647 Anti-CD4 antibody [EPR6855] ab196147 staining CD4 in Jurkat cells. The cells were fixed with 100%methanol and then incubated with 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® 647 Anti-CD4 antibody [EPR6855] ab196147 at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (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.
Anti-CD4 antibody [EPR6855] (Anti-CD4 antibody [EPR6855] ab133616)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with Anti-CD4 antibody [EPR6855] 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.
Paraffin-embedded human spleen tissue stained for CD4 using Anti-CD4 antibody [EPR6855] ab133616 at 1/500 dilution in immunohistochemical analysis.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This IHC data was generated using the same anti-CD4 antibody clone [EPR6855] in a different buffer formulation (cat# Anti-CD4 antibody [EPR6855] ab133616).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with unpurified Anti-CD4 antibody [EPR6855] ab133616 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD4 with unpurified Anti-CD4 antibody [EPR6855] ab133616.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD4 with unpurified Anti-CD4 antibody [EPR6855] ab133616.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymoma tissue labelling CD4 with unpurified Anti-CD4 antibody [EPR6855] ab133616.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Negative control: no staining on human cerebrum.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum showing no staining CD4 with purified Anti-CD4 antibody [EPR6855] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Anti-CD4 antibody [EPR6855] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD4 antibody [EPR6855] ab133616).
Tissue Microarrays stained for "Anti-CD4 antibody [EPR6855]" using "Anti-CD4 antibody [EPR6855] 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 Anti-CD4 antibody [EPR6855] 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.
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4 with 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. 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.
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4 with 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). 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.
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