Anti-BFL-1/GRS antibody [SP435]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BFL-1/GRS antibody [SP435] (AB246319)

IHC image of ab246319 staining BFL-1/GRS in a section of formalin-fixed paraffin-embedded human B-cell lymphoma* performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 30mins. The section was then incubated with ab246319, 1/117 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• WB

Lab

Western blot - Anti-BFL-1/GRS antibody [SP435] (AB246319)

Negative control :  A-673, KATO III, LoVo.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

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

All lanes:

Western blot - Anti-BFL-1/GRS antibody [SP435] (ab246319) at 1/1000 dilution

Lane 1:

KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 20 µg

Lane 2:

RKO (human colorectal Adenocarcinoma cell) whole cell lysate at 20 µg

Lane 3:

SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate at 20 µg

Lane 4:

SK-MEL-28 (human malignant melanoma cell ) at 20 µg

Lane 5:

A-673 (human muscle Ewings sarcoma cell line) whole cell lysate at 20 µg

Lane 6:

KATO III (human stomach spherical) whole cell lysate at 20 µg

Lane 7:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Observed band size: 25 kDa

false

Exposure time: 180s