Anti-BimL, clone 5E.5

Anti-BimL, clone 5E.5

Chemicon (Millipore)

Purified rat monoclonal antibody IgG2b Kappa isotype raised against recombinant GST-mouse BimL protein. The antibody recognises a reactive epitope that lies in the region that is present only in BimLong (BimL), one of the three splice variants of Bim. This epitope is absent in BimShort (BimS) and BimExtra long (BimEL). These antibodies can be used for easy detection of BimL by standard techniques including Western blot*, immunoprecipitation, immunocytochemistry, immunohistochemistry and FACS. It is highly sensitive and can, in most cases, recognise endogenous levels of BimL.

The antibody recognizes a reactive epitope that lies in the region that is present only in BimLong (BIML), one of the three splice variants of Bim. This epitope is absent in BimShort (BimS) and BimExtra Long (BimEL).

5E.5

IgG2b

• Human
• Mouse

The antibody is intended for the detection of the long splice variant of Bim in a range of techniques including:

Western blot

immunoprecipitation

FACS

immunocytochemistry

immunohistochemistry.

It should be noted that the levels of antibody quoted are intended to serve as a guide only. The optimal concentration should be determined empirically by the end user.

PROTOCOLS:

Western blot

In Western blot analysis it is recommended that the antibody be used at a concentration of 1mg/mL which has been found to detect 1x106 BimL expressing cells or 50ng of BimL recombinant protein. For the detection of endogenous BimL expression from either tissue** or cell lines**, it is recommended that the antibody be used at a concentration of 1-2mg/mL.

FACS

For FACS analysis of BimL overexpressing cell lines, it is recommended that the antibody be used at a concentration of 1-2mg/mL. It is necessary to permeabilise the cell membrane prior to analysis.

Immunoprecipitation

For immunoprecipitation of endogenous BimL from cell lines** or tissue**, it is recommended that the antibody be used at a concentration of 2mg/mL.

Immunofluorescence

In immunofluorescence techniques, it is recommended that the antibody be used at a concentration of 1-100mg/mL. The concentration should be determined empirically for each application.

*Cross reactivity to a 26kDa cytosolic protein from various kidney cell lines has been noticed in Western blots only. The identity of this protein is not known.

** Examples of cell lines tested for endogenous Bim expression are the T lymphoma cell lines B6.2.16.BW2 or KO52.DA20, B cell lymphoma line ABL5.8.1 and human breast carcinoma cell line MCF-7. It is necessary to use cell lysate from 1x107 cells. For detection of Bim expression in tissue samples, it is necessary to use 1-5mg of total protein from an appropriate tissue source.

Liquid. Antibody at a concentration of 1mg/mL containing 0.05% Bronidox as a preservative.
>95% pure as determined by SDS-PAGE.

BimL antibody is shipped at ambient temperature. Store at 2-8°C, for up to 6 months.

O43521

• NM_138622.2
• NM_138623.2
• NM_138624.2
• NM_138626.2
• NM_138627.2
• NM_207003.1
• NM_006538.3
• NM_138621.3
• NM_207002.2

• BCL2L11
• BimL
• BIM
• BIM-beta7
• BOD
• BAM
• BimEL
• BIM-beta6
• BIM-alpha6

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

The protein encoded by this gene belongs to the BCL-2 protein family. BCL-2 family members form hetero- or homodimers and act as anti- or pro-apoptotic regulators that are involved in a wide variety of cellular activities. The protein encoded by this gene contains a Bcl-2 homology domain 3 (BH3). It has been shown to interact with other members of the BCL-2 protein family, including BCL2, BCL2L1/BCL-X(L), and MCL1, and to act as an apoptotic activator. The expression of this gene can be induced by nerve growth factor (NGF), as well as by the forkhead transcription factor FKHR-L1, which suggests a role of this gene in neuronal and lymphocyte apoptosis. Transgenic studies of the mouse counterpart suggested that this gene functions as an essential initiator of apoptosis in thymocyte-negative selection. Several alternatively spliced transcript variants of this gene have been identified.

• ELISA
• Flow Cytometry
• Immunoprecipitation
• Western Blotting
• Immunohistochemistry

FUNCTION: SwissProt: O43521 # Induces apoptosis. Isoform BimL is more potent than isoform BimEL.
SIZE: 198 amino acids; 22171 Da
SUBUNIT: Forms heterodimers with a number of antiapoptotic Bcl-2 proteins including Mcl-1, Bcl-2, Bcl-X(L), BFL-1, and BHRF1. Does not heterodimerize with proapoptotic proteins such as BAD, BOK, BAX or BAK (By similarity).
SUBCELLULAR LOCATION: Membrane; Peripheral membrane protein. Note=Associated with intracytoplasmic membranes (By similarity).
DOMAIN: SwissProt: O43521 The BH3 motif is required for Bcl-2 binding and cytotoxicity.
SIMILARITY:

250 µg

Chemicon

Purified

Rat

Bim is a pro-apoptotic member of the Bcl-2 protein family. Three Bim isoforms have been reported and designated as BimEL, BimL and BimS, all of which bind Bcl2 and induce apoptosis, with BimEL being the least cytotoxic. Bim is serine phosphorylated in lymphocytes, predominantly on the EL form, which potentiates its proapoptotic activity.

Anti-BimL, clone 5E.5

Monoclonal Antibody