Name: Anti-CD4 antibody [SP35]
SKU: ab213215
Rating: 4 (3 reviews)

Anti-CD4 antibody [SP35] ab213215 is a rabbit monoclonal antibody that is used in CD4 IHC, immunofluorescence and flow cytometry. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

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Images

Flow Cytometry - Anti-CD4 antibody [SP35] (AB213215), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	mIHC	Flow Cyt	IHC-P
Human	Tested	Tested	Tested	Tested
Chimpanzee	Predicted	Predicted	Predicted	Predicted

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Chimpanzee	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Predicted
Predicted

Species	Dilution info	Notes
Species Chimpanzee	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20 - 1/100	Notes Incubate with primary antibody for 30 minutes at 4°C.

Predicted
Predicted

Species	Dilution info	Notes
Species Chimpanzee	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes Deparaffinize slides using xylene or xylene alternative and graded alcohols. Boil tissue section in EDTA buffer for 10 min followed by cooling at room temperature for 20 min. Incubate with primary antibody for 30 minutes at room temperature. Slides must be washed in between steps. Rinse slides with PBS/0.05% Tween.

Predicted
Predicted

Species	Dilution info	Notes
Species Chimpanzee	Dilution info -	Notes -

Associated Products

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Target data

See full target information CD4

Function

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:12089508, PubMed:16331979, PubMed:2214026, PubMed:9641677). Down-regulated by HIV-1 Vpu (PubMed:17346169). Acts as a receptor for Human Herpes virus 7/HHV-7 (PubMed:7909607).

Alternative names

CD4, T-cell surface glycoprotein CD4, T-cell surface antigen T4/Leu-3, CD4

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: SP35
Purification technique: Affinity purification Protein A/G
Concentration
Purification notes: Purified from TCS by protein A/G.

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

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.

11 product images

Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)
Flow cytometry analysis of THP-1 (human acute monocytic leukemia) labeling CD4 with purified ab213215 at 1/20 dilution (7.75 μg/ml) (red).
Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black). Unlableled control - Unlabeled cells (blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human Hodgkin lymphoma tissue sections labeling CD4 with ab213215 at 1/50 dilution (3.1 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human Hodgkin's lymphoma, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab213215 for 30 mins at room temperature.
Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)
Flow cytometric analysis of Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling CD4 with ab213215 at 1/100 dilution (green), compared with a rabbit IgG negative control (blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling CD4 with ab213215 at 1/50 dilution (3.1 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human tonsil, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab213215 for 30 mins at room temperature.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD4 with ab213215 at 1/50 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD4 with ab213215 at 1/50 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
Tissue Microarrays stained for "Anti-CD4 antibody [SP35]" using "ab213215"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 10 minutes. The sections were incubated with ab213215 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.
Immunocytochemistry/ Immunofluorescence - Anti-CD4 antibody [SP35] (ab213215)
Immunocytochemistry/ Immunofluorescence analysis of human PBMC (human primary peripheral blood mononuclear cell) labeling CD4 with Anti-CD4 antibody [SP35] - BSA and Azide free ab238798 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at 1/1000 dilution (green). The counterstain was Anti-human CD3 mouse monoclonal antibody (Alexa Fluor® 64) at 1/200 dilution observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% Tween-20. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)
This data was developed using Anti-CD4 antibody [SP35] - BSA and Azide free ab238798 , the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human spleen tissue labeling CD8 alpha with Anti-CD8 alpha antibody [EPR22483-288] ab245118 at 1/500 dilution, CD4 with Anti-CD4 antibody [SP35] - BSA and Azide free ab238798 at 1/500, and CD19 with Anti-CD19 antibody [SP291] - BSA and Azide free ab237772 at 1/5000 dilution.

Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human spleen.
Panel B: anti-CD8 alpha stained on cytotoxic T cells.
Panel C: anti-CD4 stained on T helper cells.
Panel D: anti-CD19 stained on B cells.

Sections were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of abab245118 for 30 mins, then Anti-CD4 antibody [SP35] - BSA and Azide free ab238798 and Anti-CD19 antibody [SP291] - BSA and Azide free ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)
This data was developed using Anti-CD4 antibody [SP35] - BSA and Azide free ab238798 , the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue labeling CD8 alpha with Anti-CD8 alpha antibody [EPR22483-288] ab245118 at 1/500 dilution, CD4 with Anti-CD4 antibody [SP35] - BSA and Azide free ab238798 at 1/500, and CD19 with Anti-CD19 antibody [SP291] - BSA and Azide free ab237772 at 1/5000 dilution.

Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B: anti-CD8 alpha stained on cytotoxic T cells.
Panel C: anti-CD4 stained on T helper cells.
Panel D: anti-CD19 stained on B cells.

Sections were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of abab245118 for 30 mins, then Anti-CD4 antibody [SP35] - BSA and Azide free ab238798 and Anti-CD19 antibody [SP291] - BSA and Azide free ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4 with ab213215 at a dilution of 1/50. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
Anti-CD4 antibody [SP35] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).