14-9887-82

antibody from Invitrogen Antibodies

Targeting: BCL6 BCL5, BCL6A, LAZ3, ZBTB27, ZNF51

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

14-9887-82

Provider

Invitrogen Antibodies

Product name

BCL6 Monoclonal Antibody (GI191E), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: This GI191E monoclonal antibody reacts with mouse and human Bcl-6, which is also known as B-cell lymphoma 6. A member of the POZ family of transcriptional repressors, Bcl-6 is expressed most abundantly in germinal center B cells, as well as in differentiating and activated B cells. This 79-kDa protein is essential for germinal center B cell proliferation and survival. Moreover, during B cell development, Bcl-6 plays an important role in promoting pre-B cell survival to enable the generation of a diverse B cell repertoire. Moreover, Bcl-6 is expressed constitutively in diffuse large B cell lymphomas (DLBCLs), Burkitt's lymphoma, and non-Hodgkin's lymphoma. Reports also suggest expression of Bcl-6 in CD4+ T follicular helper cells and memory T cells. Bcl-6 has been reported to be antagonized by Blimp-1. Applications Reported: This GI191E antibody has been reported for use in immunoblotting (WB) and immunohistochemistry (FFPE). Applications Tested: This GI191E antibody has been tested by immunoblotting of reduced cell lysate prepared from A20 cells and by immunohistochemistry on human tissue using high pH antigen retrieval. This antibody can be used at less than or equal to 5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

GI191E

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

BCL6 is critical for the development of a diverse primary B cell repertoire.

Duy C, Yu JJ, Nahar R, Swaminathan S, Kweon SM, Polo JM, Valls E, Klemm L, Shojaee S, Cerchietti L, Schuh W, Jäck HM, Hurtz C, Ramezani-Rad P, Herzog S, Jumaa H, Koeffler HP, de Alborán IM, Melnick AM, Ye BH, Müschen M
The Journal of experimental medicine 2010 Jun 7;207(6):1209-21

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Bcl6 and Blimp-1 are reciprocal and antagonistic regulators of T follicular helper cell differentiation.

Johnston RJ, Poholek AC, DiToro D, Yusuf I, Eto D, Barnett B, Dent AL, Craft J, Crotty S
Science (New York, N.Y.) 2009 Aug 21;325(5943):1006-10

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Genetic immunization: a new monoclonal antibody for the detection of BCL-6 protein in paraffin sections.

García JF, García JF, Maestre L, Lucas E, Sánchez-Verde L, Romero-Chala S, Piris MA, Roncador G
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 2006 Jan;54(1):31-8

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoblotting of reduced A20 cell lysate with 5 µg/mL of Anti-Mouse Bcl-6 Purified antibody.Bands were visualized using Anti-Mouse IgG HRP.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of Raji (Lane 1), Ramos (Lane 2), Daudi (Lane 3), Jurkat (Lane 4) and MOLT-4 (Lane 5). The blot was probed with Anti-BCL6 Monoclonal Antibody (Product # 14-9887-82, 1 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 95 kDa band corresponding to BCL6 was observed across the cell lines tested except in Jurkat and MOLT-4.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis BCL6 was performed using 70% confluent log phase Raji (B-cell lineage) cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with BCL6 Monoclonal Antibody (GI191E) (Product # 14-9887-82) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. Panel f represents negative control, MOLT-4 (T-cell lineage). The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of formalin-fixed paraffin embedded human tonsil stained with 5 µg/mL Anti-Human/Mouse Bcl-6 Purified followed by Anti-Mouse IgG Biotin, Avidin HRP, and DAB visualization.Nuclei are counterstained with hematoxylin.