Q: Please recommend which isotype antibody I should use with the NBP1-75511AF647 (Bim-AF647.) The product datasheet only says IgG, but no subclass or product number.

A: This item is a mouse IgG1 antibody. For isotype control, I would recommend NBP1-43319AF647 or NBP2-27287AF647.

Q: Would you recommend a standard Fix/Perm buffer for Bim staining?

A: Our lab used our standard ICC protocol for cell staining with this product. We used tween-20 in the permeabilization step as BIM can be found in the endomembrane system and mitochondria.

Bim Antibody

Novus Biologicals | Catalog # NBP2-53088

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Product Documents

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species Reactivity

Human, Mouse

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

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Product Specifications

Immunogen

Partial recombinant human Bim protein (amino acids 2-120) UniProt O43521]

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Bim Antibody

Western Blot: Bim Antibody [NBP2-53088]

Western Blot: Bim Antibody [NBP2-53088] - Total protein from human K562, HeLa, A549, HepG2 cells and mouse Raw264.7 cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-Bim/BCL2L11 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Immunocytochemistry/ Immunofluorescence: Bim Antibody [NBP2-53088]

Immunocytochemistry/Immunofluorescence: Bim Antibody [NBP2-53088] - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-Bim at a 1:50 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: Bim Antibody [NBP2-53088]

Immunohistochemistry-Paraffin: Bim Antibody [NBP2-53088] - IHC analysis of a formalin fixed and paraffin embedded tissue section of human endometrial cancer xenograft using Bim antibody at 1:50 dilution. The signal was detected using HRP labeled secondary antibody and DAB staining method which followed counter-staining with hematoxylin. The antibody generated a specific cytoplasmic staining with punctate appearance, signifying Bim/Bcl2L11 presence on mitochondria.

Applications for Bim Antibody

Application

Recommended Usage

Immunocytochemistry/ Immunofluorescence

10-20 ug/ml

Immunohistochemistry

1:50 - 1:100

Immunohistochemistry-Paraffin

1:50 - 1:100

Western Blot

2.0 ug/ml

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS with 1% BSA

Preservative

0.02% Sodium Azide

Concentration

0.6 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Bim

Bim (bcl-2-interacting mediator of cell death) is a proapoptotic member of the Bcl-2 family that shares only the BH3 domain with this family. There are three isoforms of Bim: BimEL, BimL, and BimS. Bim is involved in regulating the intrinsic mitochondrial apoptotic pathway by inducing cytochrome c release, which in turn, activates caspase-9 and then caspase-3. Bim also plays a critical role in central and peripheral deletion of T lymphocytes and in controlling B cell homeostasis and activation. BimEL, the long isoform of Bim, is a ~28 kDa protein that is predominantly expressed in T and B cells and is activated by ERK1/2 pathway. The activation of BimEL by ERK1/2 promotes its phosphorylation on serine 65 (serine 69 in human), targeting it for degradation via the proteasome.

Alternate Names

BAM, bcl-2 interacting mediator of cell death, bcl-2 interacting protein Bim, Bcl2-interacting mediator of cell death, Bcl2-L-11, BCL2-like 11 (apoptosis facilitator), bcl-2-like protein 11, bcl-2-related ovarian death agonist, BIM-alpha6, BIM-beta6, BIM-beta7, BIMBOD, BimEL, BimL

Gene Symbol

BCL2L11

UniProt

O43521 (Human)

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Product Documents for Bim Antibody

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Certificate of Analysis

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Product Specific Notices for Bim Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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