PA5-27390

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-27390

Provider

Invitrogen Antibodies

Product name

BCL6 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: Jurkat, K562, THP-1, Neuro2A, GL261, C8D30, NIH-3T3, Raw264.7, C2C12. Predicted reactivity: Mouse (91%), Rat (91%), Dog (90%), Pig (89%), Rhesus Monkey (97%), Bovine (90%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1.27 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of BCL6 using 30 µg of A) Jurkat (B) Raji and C) NCI-H929 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a BCL6 polyclonal antibody (Product # PA5-27390) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of BCL6 using A) 30 µg Neuro2A whole cell lysate (B) 30 µg GL261 whole cell lysate (C) 30 µg C8D30 whole cell lysate (D) 30 µg NIH-3T3 whole cell lysate (E) 30 µg Raw264.7 whole cell lysate and F) 30 µg C2C12 whole cell lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a BCL6 polyclonal antibody (Product # PA5-27390) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of BCL6 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS PAGE. Proteins were transferred to a membrane and probed with a BCL6 Polyclonal Antibody (Product # PA5-27390) at a dilution of 1:1000. A: Jurkat, B: K562.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using BCL6 Polyclonal Antibody (Product # PA5-27390). Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with BCL6 Polyclonal Antibody (Product # PA5-27390) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

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 Polyclonal Antibody (Product # PA5-27390) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 (Frozen) analysis of BCL6 was performed in frozen-sectioned mouse cerebellum tissue using BCL6 Polyclonal Antibody (Product # PA5-27390) at a dilution of 1:250 (Green). Red: NF-H, stained by NF-H antibody diluted at 1:500. Blue: Fluoroshield with DAPI.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Cross-linked ChIP was performed with Raji chromatin extract and 5 µg of either control rabbit IgG or BCL6 Polyclonal Antibody (Product # PA5-27390). The precipitated DNA was detected by PCR with primer set targeting to PRDM1 or CD20.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

BCL6 antibody immunoprecipitates BCL6 protein in IP experiments. IP Sample: Raji whole cell lysate/extract A : 30 µg whole cell lysate/extract of BCL6 protein expressing Raji cells B : Control with 3 µg of pre-immune rabbit IgG C : Immunoprecipitation of BCL6 by 3 µg of BCL6 antibody (Product # PA5-27390) 7.5% SDS-PAGE The immunoprecipitated BCL6 protein was detected by BCL6 antibody (Product # PA5-27390) diluted at 1:1,000. Anti-rabbit IgG (HRP) was used as a secondary reagent.