Invitrogen

BID Monoclonal Antibody (23F7)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. View Details

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Datasheet

Questions & Answers

Datasheet

Questions & Answers

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 was performed using Anti-BID Monoclonal Antibody (23F7) (Product # MA1-13043) and a 21 kDa band corresponding to BID was observed across cell lines tested and decreased upon Staurosporine treatment. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), A-431 treated with Staurosporine (1µM for 8 hour) (Lane 2), MCF-7 (Lane 3) and MCF-7 treated with Staurosporine (1µM for 8 hour) (Lane 4) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

BID Antibody in Cell treatment — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using BID Monoclonal Antibody (23F7) (Product # MA1-13043), shows decreased expression of proteins in both A-431 and MCF-7 upon Staurosporine treatment. {TM}

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

1-2 µg/mL

Immunofluorescence (IF)

1-2 µg/mL

Western Blot (WB)

1:1000

Product Specifications

Species

Human, Mouse

Expression System

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.

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Bioinformatics

Protein Aliases: apoptic d; 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; HGNC:1050; Human BID coding sequence; MGC15319; MGC42355; OTTHUMP00000196197; 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)