Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free
See all IRF5 primary antibodies
Description

Rabbit monoclonal [EPR23312-91] to IRF5 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Mouse
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: THP-1, A20, NR8383, Ramos and RAW 264.7 whole cell lysates: Mouse spleen, thymus, and lymph node tissue lysates; Rat lymph node and thymus tissue lysates. ICC: A20 and THP-1 cells. Flow cyt: A20 and THP-1 cells. IP: A20 and THP-1 whole cell lysates.
General notes
ab274379 is the carrier-free version of ab274373. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm.

Maxpar^® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23312-91
Isotype

IgG
Research areas

Western blot - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

All lanes : Anti-IRF5 antibody [EPR23312-91] (ab274373) at 1/1000 dilution

Lanes 1 & 5 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lanes 2 & 7 : A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate
Lane 3 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 4 : MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate
Lane 6 : Ramos (human burkitts lymphoma b lymphocyte) whole cell lysate
Lane 8 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 56 kDa
Observed band size: 56 kDa

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

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

Lysates loaded onto lanes 1 and 2 were made freshly and used in WB immediately to minimize protein degradation. Bands beneath the target band in lanes 5 and 7-8 may be caused by degradation.

Negative control: K-562 and MOLT-4 (PMID: 11303025).

The blot of lanes 5-8 was developed using a higher sensitivity ECL substrate.

Exposure time: 3 minutes.
Immunocytochemistry - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (Human monocytic leukemia monocyte) cells labelling IRF5 with ab274373 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (Green). Confocal image showing strong cytoplasmic and weak nuclear staining in THP-1 cells is observed.

Tubulin is detected with Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized THP-1 (Human monocytic leukemia cell line) cells labelling IRF5 with ab274373 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG isotype control (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

IRF5 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate with ab274373 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274373 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: THP-1 whole cell lysate 10 ug.

Lane 2: ab274373 IP in THP-1 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab274373 in THP-1 whole cell lysate

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

Exposure time: 15 seconds.

Lysate was made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.
Western blot - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

All lanes : Anti-IRF5 antibody [EPR23312-91] (ab274373) at 1/1000 dilution

Lane 1 : Mouse spleen tissue lysate
Lane 2 : Mouse thymus tissue lysate
Lane 3 : Mouse lymph node tissue lysate
Lane 4 : Rat thymus tissue lysate
Lane 5 : Rat lymph node tissue lysate
Lane 6 : NR8383 (rat norvegicus lu macrophage (alveolar)) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 56 kDa
Observed band size: 56 kDa

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

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

Bands beneath the target band may be caused by degradation.

The blot was developed using a higher sensitivity ECL substrate.

Exposure time: 3 minutes.
Immunocytochemistry - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A20 (Mouse reticulum sarcoma B lymphocyte) cells labelling IRF5 with ab274373 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (Green). Confocal image showing strong cytoplasmic and weak nuclear staining in A20 cells is observed.

Tubulin is detected with Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized A20 (Mouse reticulum sarcoma B lymphocyte) cells labelling IRF5 with ab274373 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG isotype control (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)

This data was developed using ab274373, the same antibody clone in a different buffer formulation.

IRF5 was immunoprecipitated from 0.35 mg A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate with ab274373 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274373 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: A20 whole cell lysate 10 ug.

Lane 2: ab274373 IP in A20 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab274373 in A20 whole cell lysate.

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

Exposure time: 15 seconds.

Lysate was made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.
Anti-IRF5 antibody [EPR23312-91] - BSA and Azide free (ab274379)