Invitrogen

BID Monoclonal Antibody (23F7)

Product Image Gallery

BID Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of BID was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with BID (23F7) Mouse Monoclonal Antibody (Product # MA1-13043) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature 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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300).Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

BID Antibody in Western Blot (WB) — Western blot analysis of BID was performed by loading 0.125-1 µg purified BID-GST fusion protein onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/0.1%TBST for at least 1 hour. Membranes were then probed with mouse monoclonal BID clone 23F7 antibody (Product # MA1-13043) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS/0.1%Tween-20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 31430) at a dilution of 1:25000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Pierce Super Signal West Dura (Product # 34075).

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

1-2 µg/mL

Immunofluorescence (IF)

1-2 µg/mL

Western Blot (WB)

1:1000

Product Specifications

Species Reactivity

Human, Mouse

Host / Isotype

Mouse / IgG

Class

Monoclonal

Type

Antibody

Clone

23F7

Immunogen

Residues 5-195 from human recombinant protein expressed in bacteria

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

PBS with 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_10985736

Product Specific Information

MA1-13043 detects BID from human and mouse samples.

MA1-13043 has been successfully used in ICC/IF and Western blot procedures. By Western blot, this antibody detects a 22 kDa protein corresponding to human BID.

The MA1-13043 antigen is a recombinant protein corresponding to residues 5-195 of human BID.

Target Information

BID is a death agonist that heterodimerizes with either agonist BAX or antagonist BCL2. Bid is a member of the BCL-2 family of cell death regulators. BID is a mediator of mitochondrial damage induced by caspase-8 (CASP8); CASP8 cleaves this encoded protein, and the COOH-terminal part translocates to mitochondria where it triggers cytochrome c release. BID is also a pro-apoptotic member of the Bcl 2 family. BID usually exists in an inactive form in the cytosolic fraction of living cells and becomes cleaved and activated by caspase 8 in response to TNF alpha or Fas ligand. Once BID is cleaved, the C-terminal 15 kDa fragment of BID (p15) translocates onto mitochondria and is sufficient to trigger cytochrome c release, resulting in cell apoptosis. BID serves as a direct molecular link between caspase 8 activation and mitochondrial death machinery.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: apoptic death agonist; BH3-interacting domain death agonist; BH3-interacting domain death agonist p11; BH3-interacting domain death agonist p13; BH3-interacting domain death agonist p15; BID; BID isoform ES(1b); BID isoform L(2); BID isoform Si6; desmocollin type 4; Human BID coding sequence; p11 BID; p13 BID; p15 BID; p22 BID

Gene Aliases: 2700049M22Rik; AI875481; AU022477; BID; FP497

UniProt ID: (Human) P55957, (Mouse) P70444

Entrez Gene ID: (Human) 637, (Mouse) 12122

Function(s)