Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Plant - Arabidopsis thaliana

Applications

Validated:

Western Blot, Simple Western, Immunoprecipitation

Cited:

Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

E. coli-derived recombinant mouse BID
Met1-Asp195
Accession # AAC71064

Specificity

Detects human and mouse BID, a fragment generated in cells treated with anti-Fas and the carboxyl‑terminal portion of recombinant BID generated by cleavage with Caspase 8 in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse BID Antibody

Detection of Human BID antibody by Western Blot.

Detection of Human BID by Western Blot.

Western blot shows Jurkat human acute T cell leukemia cell line treated with apoptosis inducer anti-Fas for the indicated times. PVDF membrane was probed with 1 µg/mL Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BID at approximately 20 kDa (as indicated). For additional reference short (5 seconds, left panel) and long (60 seconds, right panel) exposures are shown. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Human and Mouse BID antibody by Western Blot.

Detection of Human and Mouse BID by Western Blot.

Western blot shows lysates of SVEC4-10 mouse vascular endothelial cell line and Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for BID at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human BID antibody by Simple WesternTM.

Detection of Human BID by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for BID at approximately 29 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human BID by Western Blot

Detection of Human BID by Western Blot

Survivin is predominantly expressed in obese hASCs and is influenced by the inflammatory environment. (a) Survivin protein expression and quantification in total adipose tissue (AT), adipocyte fraction (AD) and stromal-vascular fraction (SVF) from SAT of lean and obese patients. Fumarylacetoacetase (FAA) was used as a loading control. Ponceau S staining is also provided. *P<0.001 versus total AT and AD. (b) Survivin and p53 protein levels and quantification in hASCs from lean and obese patients. GAPDH was used as a loading control. *P<0.01 versus lean. (c) Survivin secreted levels and quantification in medium from lean- and obese-derived hASCs. Ponceau S staining was used to check loading. *P<0.05 versus lean conditioned medium. (d) Total cell extracts of lean- and obese-derived hASCs were subjected to immunoblotting with antibodies against apoptotic and antiapoptotic markers and GAPDH was used as a loading control. No significant differences were found. (e) Lean hASCs were cultured in RPMI medium (C: control) or for 24 h in conditioned medium (CM) from macrophages (M0, M1 and M2) and cells were subjected to immunobloting with survivin and GAPDH antibodies. *P<0.05 versus CM from control, and M0; #P<0.05 versus M2. (f) Left panel, lean hASCs were cultured in RPMI medium (C: control) or RPMI supplemented with IL-1 beta (IL1B) or IL-1 beta and IL-1 beta receptor antagonist (a); right panel, with CM of M1 macrophages or CM of M1 and IL-1 beta receptor antagonist (a). Survivin protein levels were measured and GAPDH was used as a loading control. Quantification of three experiments is shown. *P<0.05 versus control (c); #P<0.05 as indicated in figure. Data information: Values are expressed as mean±S.E.M. Statistical analysis: unpaired Student’s t-test. n=3–4 patients for each group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28518147), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human BID by Western Blot

Detection of Human BID by Western Blot

Survivin is predominantly expressed in obese hASCs and is influenced by the inflammatory environment. (a) Survivin protein expression and quantification in total adipose tissue (AT), adipocyte fraction (AD) and stromal-vascular fraction (SVF) from SAT of lean and obese patients. Fumarylacetoacetase (FAA) was used as a loading control. Ponceau S staining is also provided. *P<0.001 versus total AT and AD. (b) Survivin and p53 protein levels and quantification in hASCs from lean and obese patients. GAPDH was used as a loading control. *P<0.01 versus lean. (c) Survivin secreted levels and quantification in medium from lean- and obese-derived hASCs. Ponceau S staining was used to check loading. *P<0.05 versus lean conditioned medium. (d) Total cell extracts of lean- and obese-derived hASCs were subjected to immunoblotting with antibodies against apoptotic and antiapoptotic markers and GAPDH was used as a loading control. No significant differences were found. (e) Lean hASCs were cultured in RPMI medium (C: control) or for 24 h in conditioned medium (CM) from macrophages (M0, M1 and M2) and cells were subjected to immunobloting with survivin and GAPDH antibodies. *P<0.05 versus CM from control, and M0; #P<0.05 versus M2. (f) Left panel, lean hASCs were cultured in RPMI medium (C: control) or RPMI supplemented with IL-1 beta (IL1B) or IL-1 beta and IL-1 beta receptor antagonist (a); right panel, with CM of M1 macrophages or CM of M1 and IL-1 beta receptor antagonist (a). Survivin protein levels were measured and GAPDH was used as a loading control. Quantification of three experiments is shown. *P<0.05 versus control (c); #P<0.05 as indicated in figure. Data information: Values are expressed as mean±S.E.M. Statistical analysis: unpaired Student’s t-test. n=3–4 patients for each group Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28518147), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse BID Antibody

Application
Recommended Usage

Immunoprecipitation

4 µg/106 cells
Sample: Jurkat human acute T cell leukemia cell line treated with anti-Fas, see our available Western blot detection antibodies

Simple Western

10 µg/mL
Sample: Jurkat human acute T cell leukemia cell line

Western Blot

1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line was treated or untreated with anti-Fas and SVEC4‑10 mouse vascular endothelial cell line

Reviewed Applications

Read 4 reviews rated 4 using AF860 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: BID

BID is a 195 amino acid member of the Bcl-2 family of proteins that regulates outer mitochondrial membrane permeability (1). BID is a pro-apoptotic member that causes cytochrome c to be released from the mitochondria intermembrane space into the cytosol. In healthy cells BID is cytosolic. In response to Fas ligand or TNF, BID is cleaved by caspase-8 and it then relocates to the mitochondria outer membrane (2, 3). Cleavage of BID by caspase-8 generates a new N-terminal that contains a terminal glycine. It appears that the glycine is myristoylated and myristoylation serves to target BID to the mitochondria (4). BID may then interact with another pro-apoptotic Bcl-2 family member Bak (5). Interaction of BID with Bak causes altered mitochondrial membrane permeability. A 9‑13 amino acid stretch called the BH3 region (Bcl-2 homology region) appears to mediate the BID interaction with other Bcl-2 family members. BID is neutralized by binding to the anti-apoptotic member Bcl-xL.

References

  1. Gross, A. et al. (1999) Genes and Develop. 13:1899.
  2. Luo, X., et al. (1998) Cell 94:481.
  3. Li, H. et al. (1998) Cell 94:491.
  4. Zha, J. et al. (2000) Science 290:1761.
  5. Wei, M.C. et al. (2000) Genes Dev. 14:2060.

Long Name

BH3 Interacting Domain Death Agonist

Alternate Names

apoptic death agonist, BH3 interacting domain death agonist, BH3-interacting domain death agonist, BID isoform ES(1b), BID isoform L(2), BID isoform Si6, desmocollin type 4, FP497, Human BID coding sequence, MGC15319, MGC42355, p22 BID, tbid

Entrez Gene IDs

637 (Human); 12122 (Mouse)

Gene Symbol

BID

UniProt

Additional BID Products

Product Documents for Human/Mouse BID Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse BID Antibody

For research use only

Related Research Areas

Citations for Human/Mouse BID Antibody

Customer Reviews for Human/Mouse BID Antibody (4)

4 out of 5
4 Customer Ratings
5 Stars
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4 Stars
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3 Stars
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2 Stars
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1 Stars
0%

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Showing  1 - 4 of 4 reviews Showing All
Filter By:
  • Human/Mouse BID Antibody
    Name: Anonymous
    Application: Microarrays
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 03/11/2019
    Human/Mouse BID Antibody AF860
  • Human/Mouse BID Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Plasma
    Species: Human
    Verified Customer | Posted 11/10/2018
  • Human/Mouse BID Antibody
    Name: Carly Garrison
    Application: Microarray
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 11/09/2018
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 21048194
    Species: Human
    Verified Customer | Posted 01/09/2015

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