Anti-IRF4 antibody [EPR5653]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF4 antibody [EPR5653] (AB124691)

ab124691, at 1/100, staining IRF4 in paraffin-embedded Human colon tissue by Immunohistochemistry.

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF4 antibody [EPR5653] (AB124691)

ab124691, at 1/100, staining IRF4 in paraffin-embedded Human tonsil tissue by Immunohistochemistry.

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

• ChIP

Lab

ChIP - Anti-IRF4 antibody [EPR5653] (AB124691)

Chromatin was prepared from SK-MEL-28 cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 μg of chromatin, 5 μg of ab124691 (red), or 5 μg of rabbit normal IgG ab172730 (gray) and 25 μl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

ChIP was performed according to the literature (PMID : 24267888)

• IP

Lab

Immunoprecipitation - Anti-IRF4 antibody [EPR5653] (AB124691)

IRF4 was immunoprecipitated from 0.35 mg Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 μg with 124691 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 μg

Lane 2 : ab124691 IP in Ramos whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab124691 in Ramos whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-IRF4 antibody [EPR5653] (ab124691)

Predicted band size: 52 kDa

Observed band size: 52 kDa

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• WB

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Western blot - Anti-IRF4 antibody [EPR5653] (AB124691)

All lanes:

Western blot - Anti-IRF4 antibody [EPR5653] (ab124691) at 1/1000 dilution

Lane 1:

Ramos cell lysate at 10 µg

Lane 2:

HuT-78 cell lysate at 10 µg

Lane 3:

IM 9 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated at 1/2000 dilution

Predicted band size: 52 kDa

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• IHC-P

AbReview33323****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF4 antibody [EPR5653] (AB124691)

Immunohistochemical analysis of dog spleen tissue, staining IRF4 with ab124691.

Tissue was fixed with formaldehyde and blocked with 2.5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a TRIS-EDTA buffer (pH 9). Samples were incubated with primary antibody (1/50 in PBS plus 1X casein) for 1 hour 30 minutes at 37°C. An undiluted HRP-conjugated horse anti-rabbit polyclonal IgG was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• OI-RD Scanning

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OI-RD Scanning - Anti-IRF4 antibody [EPR5653] (AB124691)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.