Anti-CD4 antibody [EPR19514]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD4 with ab183685 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on T cells is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

AbReview63464****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formalin-fixed 0.2% Triton-X permeabilized mouse Eo771 mammary tumor tissue stained for CD4 with ab183685 (18 hours 4°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG (H&L) polyclonal AlexaFluor^®488 conjugate was used as the secondary (green).

Heat mediated antigen retrieval buffer/enzyme used : Citrate buffer.

Blocking step : 4% BSA for 1 hour at 25°C.

Anugraha Rajagopalan

• IHC-P

AbReview68740****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed non-permeabilized mouse spleen tissue stained for CD4 with ab183685 (30mins at a 1/1000 dilution) in immunohistochemical analysis. A Goat polyclonal HRP conjugate was used as the secondary.

Heat mediated antigen retrieval buffer/enzyme used : Leica ER2 solution.

Zohar Gavish

• IHC-P

AbReview58971****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed paraffin-embedded mouse spleen tissue stained for CD4 using ab183685 at 1/2000 dilution in immunohistochemical analysis followed by Goat anti Rabbit IgG Biotin.

This image is courtesy of an Abreview submitted by Mr. Carl Hobbs.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD4 with ab183685 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative on mouse cerebrum.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

AbReview68614****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed 3% hydrogen peroxide permeabilized B16F10 (melanoma) syngeneic mouse tumor tissue stained for CD4 with ab183685 (36mins 37°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG polyclonal HRP conjugate was used as the secondary.

Heat mediated antigen retrieval buffer/enzyme used : Tris/EDTA.

Verified customer

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling CD4 with ab183685 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on lymphocytes and negative on epithelium cells of mouse colon is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

AbReview59119****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed mouse colon chronic mucosal inflammation tissue stained for CD4 with ab183685 (90 mins at a 1/1000 dilution) in immunohistochemical analysis. A Goat polyclonal biotin conjugate was used as the secondary.

Heat mediated antigen retrieval buffer/enzyme used : High pH CC1.

Blocking step : 1% BSA for 40 mins at 24°C.

Verified customer

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Immunohistochemical analysis of 4% paraformaldehyde-fixed 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling CD4 with ab183685 at 1/500 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

The result showed membrane staining on mouse spleen.

The nuclear counterstain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab150077 at 1/1000 dilution.

Antigen retrieval : Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

AbReview66989****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed paraffin-embedded mouse lung tissue stained for CD4 with ab183685 (18 hours 4°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG polyclonal HRP conjugate was used as the secondary at a 1/250 dilution (red).

Heat mediated antigen retrieval buffer/enzyme used : Citrate buffer pH 6.0.

Blocking step : 10% serum for 1 hour at 25°C.

Verified customer

• IHC-Fr

AbReview61721****

Immunohistochemistry (Frozen sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Paraformaldehyde-fixed 0.1% Tween 0.3% Triton in PBS permeabilized mouse liver tissue stained for CD4 with ab183685 (12 hours 4°C at a 1/100 dilution) in immunohistochemical analysis. A Donkey anti Rabbit IgG (H+L) AlexaFluor^®647 was used as the secondary at a 1/500 dilution (red).

Blocking step : 1% BSA for 12 hours at 4°C.

Verified customer

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-CD4 antibody [EPR19514] (AB183685)

IHC image of CD4 staining in a section of frozen normal Mouse Spleen.

The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab183685 (Rabbit monoclonal [EPR19514] to CD4) at 1/200 and ab243840 at 1μg/ml to show the distinct staining of B cells and T cells. The section was then incubated with ab150165 (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preabsorbed (Shown in green) 1/1000) and ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor®594) (Shown in red) 1/1000) for 1 hour at room temperature. The secondary-only control insert image is taken from an identical assay without primary antibody. DAPI was used to stain the cell nuclei (blue). The section was then mounted using Fluoromount®.

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

For IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antibody concentrations and incubation times.

• IHC-P

AbReview64045****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed paraffin-embedded mouse thymus tissue stained for CD4 using ab183685 at 1/500 dilution in immunohistochemical analysis.

This image is courtesy of an anonymous Abreview.

• IHC-P

AbReview62644****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed non-permeabilized mouse colon tissue stained for CD4 with ab183685 (30 mins at a 1/500 dilution) in immunohistochemical analysis. A Rabbit polyclonal HRP conjugate was used as the secondary.

Heat mediated antigen retrieval buffer/enzyme used : pH 9.0 EDTA.

Blocking step : 1% ab64226 for 10 mins at RT.

Verified customer

• IHC-P

AbReview57043****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

10% NBF-fixed paraffin-embedded mouse spleen tissue stained for CD4 using ab183685 at 1/2000 dilution in immunohistochemical analysis followed by Goat anti-Rabbit IgG Alexa Fluor^® 647.

This image is courtesy of an anonymous Abreview.

• IHC-P

AbReview65667****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (AB183685)

Formaldehyde-fixed non-permeabilized mouse brain tissue stained for CD4 with ab183685 (14 hours 4°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG monoclonal HRP conjugate was used as the secondary at a 1/200 dilution.

Blocking step : 5% normal goat serum for 1 hour at 26°C.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Herr Dr. Markus Kipp

• IP

Supplier Data

Immunoprecipitation - Anti-CD4 antibody [EPR19514] (AB183685)

CD4 was immunoprecipitated from 1mg of Mouse thymus whole cell lysate with ab183685 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab183685 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Mouse thymus whole cell lysate, 10μg (Input).

Lane 2 : ab183685 IP in Mouse thymus whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab183685 in Mouse thymus whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-CD4 antibody [EPR19514] (ab183685)

Predicted band size: 51 kDa

false

• WB

Lab

Western blot - Anti-CD4 antibody [EPR19514] (AB183685)

The expression profile observed is consistent with what has been described in the literature (PMID : 2155425).

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 3 seconds; Lanes 2,3,4,5 and 6 : 30 seconds

All lanes:

Western blot - Anti-CD4 antibody [EPR19514] (ab183685) at 1/2000 dilution

Lane 1:

Mouse thymus lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate at 20 µg

Lane 6:

NIH/3T3 (mouse embryo) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 51 kDa

Observed band size: 51 kDa

false