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    bag1-antibody-y166-ab32109.pdf

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Cell Biology Apoptosis Intracellular Bcl2 Family
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RecombinantRabMAb

Recombinant Anti-Bag1 antibody [Y166] (ab32109)

  • Datasheet
Reviews (4) Submit a question References (8)

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Western blot - Anti-Bag1 antibody [Y166] (ab32109)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bag1 antibody [Y166] (ab32109)
  • Immunocytochemistry/ Immunofluorescence - Anti-Bag1 antibody [Y166] (ab32109)
  • Anti-Bag1 antibody [Y166] (ab32109)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y166] to Bag1
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Human

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Overview

  • Product name

    Anti-Bag1 antibody [Y166]
    See all Bag1 primary antibodies
  • Description

    Rabbit monoclonal [Y166] to Bag1
  • Host species

    Rabbit
  • Specificity

    The antibody should recognize all three isoforms of Bag-1 (Bag-1L, Bag-1M and Bag-1S).
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human Bag1 aa 50-150. The exact sequence is proprietary.

  • Positive control

    • HeLa cell lysate and cells, human prostate carcinoma tissue
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y166
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Bcl2 Family
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Bcl 2 family
    • Cardiovascular
    • Atherosclerosis
    • Apoptosis
    • Cardiovascular
    • Heart
    • Autophagy
    • Autophagosome
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Autophagosome
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Bcl 2 family
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Cancer
    • Cell Death
    • Autophagy
    • Autophagosome

Associated products

  • Alternative Versions

    • Anti-Bag1 antibody [Y166] - BSA and Azide free (ab211754)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant Human Bag1 protein (ab85158)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32109 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (2)
1/500. Detects a band of approximately 33, 46, 50 kDa (predicted molecular weight: 31 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF (1)
1/100 - 1/250.
Notes
WB
1/500. Detects a band of approximately 33, 46, 50 kDa (predicted molecular weight: 31 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
Application notes
Is unsuitable for Flow Cyt.

Target

  • Function

    Inhibits the chaperone activity of HSP70/HSC70 by promoting substrate release. Inhibits the pro-apoptotic function of PPP1R15A, and has anti-apoptotic activity. Markedly increases the anti-cell death function of BCL2 induced by various stimuli.
  • Tissue specificity

    Isoform 4 is the most abundantly expressed isoform. It is ubiquitously expressed throughout most tissues, except the liver, colon, breast and uterine myometrium. Isoform 1 is expressed in the ovary and testis. Isoform 4 is expressed in several types of tumor cell lines, and at consistently high levels in leukemia and lymphoma cell lines. Isoform 1 is expressed in the prostate, breast and leukemia cell lines. Isoform 3 is the least abundant isoform in tumor cell lines (at protein level).
  • Sequence similarities

    Contains 1 BAG domain.
    Contains 1 ubiquitin-like domain.
  • Post-translational
    modifications

    Ubiquitinated; mediated by SIAH1 or SIAH2 and leading to its subsequent proteasomal degradation.
  • Cellular localization

    Cytoplasm. Nucleus. Isoform 2 localizes to the cytoplasm and shuttles into the nucleus in response to heat shock; Nucleus. Cytoplasm. Isoform 1 localizes predominantly to the nucleus and Cytoplasm. Nucleus. Isoform 4 localizes predominantly to the cytoplasm. The cellular background in which it is expressed can influence whether it resides primarily in the cytoplasm or is also found in the nucleus. In the presence of BCL2, localizes to intracellular membranes (what appears to be the nuclear envelope and perinuclear membranes) as well as punctate cytosolic structures suggestive of mitochondria.
  • Target information above from: UniProt accession Q99933 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 573 Human
    • Entrez Gene: 12017 Mouse
    • Entrez Gene: 297994 Rat
    • Omim: 601497 Human
    • SwissProt: Q99933 Human
    • SwissProt: Q60739 Mouse
    • SwissProt: B0K019 Rat
    • Unigene: 377484 Human
    • Unigene: 688 Mouse
    • Unigene: 16320 Rat
    see all
  • Alternative names

    • Bag 1 antibody
    • BAG family molecular chaperone regulator 1 antibody
    • BAG-1 antibody
    • BAG1 antibody
    • BAG1_HUMAN antibody
    • BCL 2 Associated Athanogene 1 antibody
    • BCL 2 associated athanogene antibody
    • BCL 2 binding athanogene 1 antibody
    • Bcl-2-associated athanogene 1 antibody
    • BCL2 associated athanogene 1 antibody
    • BCL2 associated athanogene antibody
    • Glucocorticoid receptor-associated protein RAP46 antibody
    • HAP1 antibody
    • Haptoglobin antibody
    • RAP 46 antibody
    • RAP46 antibody
    see all

Images

  • Western blot - Anti-Bag1 antibody [Y166] (ab32109)
    Western blot - Anti-Bag1 antibody [Y166] (ab32109)
    Anti-Bag1 antibody [Y166] (ab32109) at 1/500 dilution + HeLa cell lysate

    Predicted band size: 31 kDa
    Observed band size: 33 kDa why is the actual band size different from the predicted?
    Additional bands at: 46 kDa (possible isoform), 50 kDa (possible isoform)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bag1 antibody [Y166] (ab32109)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bag1 antibody [Y166] (ab32109)

    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using ab32109 at 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Bag1 antibody [Y166] (ab32109)
    Immunocytochemistry/ Immunofluorescence - Anti-Bag1 antibody [Y166] (ab32109)
    Immunofluorescent staining of HeLa cells using ab32109 at 1/100 dilution.
  • Anti-Bag1 antibody [Y166] (ab32109)
    Anti-Bag1 antibody [Y166] (ab32109)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (8)

Publishing research using ab32109? Please let us know so that we can cite the reference in this datasheet.

ab32109 has been referenced in 8 publications.

  • Gennaro VJ  et al. Interaction between the BAG1S isoform and HSP70 mediates the stability of anti-apoptotic proteins and the survival of osteosarcoma cells expressing oncogenic MYC. BMC Cancer 19:258 (2019). PubMed: 30902071
  • Miao JT  et al. LncRNA ANRIL affects the sensitivity of ovarian cancer to cisplatin via regulation of let-7a/HMGA2 axis. Biosci Rep 39:N/A (2019). PubMed: 31189742
  • Lu S  et al. Downregulation of BAG-1 in T47D cells promotes resistance to tamoxifen via activation of the PI3K/Akt/mTOR signaling pathway. Oncol Rep 41:1901-1910 (2019). PubMed: 30747221
  • Shen Z  et al. miR-494-BAG-1 axis is involved in cinobufacini-induced cell proliferation and apoptosis in gastric cancer. Mol Med Rep 17:7435-7441 (2018). PubMed: 29568970
  • Wood RJ  et al. A biosensor-based framework to measure latent proteostasis capacity. Nat Commun 9:287 (2018). PubMed: 29348634
  • Cato L  et al. Development of Bag-1L as a therapeutic target in androgen receptor-dependent prostate cancer. Elife 6:N/A (2017). IHC-P, WB ; Human . PubMed: 28826504
  • Mashukova A  et al. The BAG-1 isoform BAG-1M regulates keratin-associated Hsp70 chaperoning of aPKC in intestinal cells during activation of inflammatory signaling. J Cell Sci 127:3568-77 (2014). PubMed: 24876225
  • Anderson LR  et al. BAG-1 overexpression attenuates luminal apoptosis in MCF-10A mammary epithelial cells through enhanced RAF-1 activation. Oncogene 29:527-38 (2010). ICC/IF ; Human . PubMed: 19881545

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Bag1 antibody [Y166]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Sample
Human Cell (Human primary fibroblasts)
Specification
Human primary fibroblasts
Permeabilization
Yes - 0.2% Triton-X100
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 11 2013

Western blot abreview for Anti-Bag1 antibody [Y166]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Loading amount
20 µg
Specification
Mouse embryonic fibroblasts
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly gel)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 23 2011

Immunoprecipitation abreview for Anti-Bag1 antibody [Y166]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa cells)
Total protein in input
200 µg
Specification
HeLa cells
Immuno-precipitation step
Protein A/G
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 06 2011

Western blot abreview for Anti-Bag1 antibody [Y166]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Loading amount
20 µg
Specification
HeLa cells
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly gel)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Svetlana Khoronenkova

Verified customer

Submitted Aug 25 2010

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