39-8800

IRF8 antibody from Invitrogen Antibodies

ICSBP, ICSBP1, IRF-8

Western blot

Supportive data in Antibodypedia

ELISA

Recommended by provider

Flow cytometry

Supportive data in Antibodypedia

Provider product page for 39-8800

Antibody Data

Product number

39-8800

Provider

Invitrogen Antibodies

Product name

Anti-IRF8 Monoclonal Antibody (ZI003)

Provider product page

Antibody type

Monoclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

ZI003

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

-20°C

References

Identification of IRF8 as a potent tumor suppressor in murine acute promyelocytic leukemia.
Gaillard C, Surianarayanan S, Bentley T, Warr MR, Fitch B, Geng H, Passegué E, de Thé H, Kogan SC
Blood advances 2018 Oct 9;2(19):2462-2466

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Ramos cell lysates using Zymed Ms anti-ICSBP (Product # 39-8800).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on nuclear enriched lysates (30 µg lysate) of Raji (Lane 1), Ramos (Lane 2), and KARPAS-299 (Lane 3). The blots were probed with Anti-IRF8 Mouse Monoclonal Antibody (Product # 39-8800, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 48 kDa band corresponding to IRF8 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on nuclear enriched extracts (30 µg lysate) of Raji (Lane 1), Ramos (Lane 2), K-562 (Lane 3), HL-60 (Lane 4), U-2 OS (Lane 5),THP-1 (Lane 6), NIH/3T3 (Lane 7) and T98G (Lane 8). The blot was probed with Anti- IRF8 Mouse Monoclonal Antibody (Product # 39-8800, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 48 kDa band corresponding to IRF8 was observed in Raji, Ramos and not observed in other cell lines which are documented to be IRF8 negative.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on nuclear enriched extracts (30 µg lysate) of Raji (Lane 1), Ramos (Lane 2), K-562 (Lane 3), HL-60 (Lane 4), U-2 OS (Lane 5),THP-1 (Lane 6), NIH/3T3 (Lane 7) and T98G (Lane 8). The blot was probed with Anti- IRF8 Mouse Monoclonal Antibody (Product # 39-8800, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 48 kDa band corresponding to IRF8 was observed in Raji, Ramos and not observed in other cell lines which are documented to be IRF8 negative.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of IRF8 was done on U-937 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with IRF8 Mouse Monoclonal Antibody (398800, red histogram) or with mouse isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control..