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Validated using a knockout cell line

Anti-Bcl-2 antibody [100/D5] (ab692)

Reviews (9)Q&A (7)References (211)
Western blot - Anti-Bcl-2 antibody [100/D5] (ab692)
  • Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 antibody [100/D5] (ab692)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [100/D5] (ab692)
  • Flow Cytometry - Anti-Bcl-2 antibody [100/D5] (ab692)

Key features and details

  • Mouse monoclonal [100/D5] to Bcl-2
  • Suitable for: WB, Flow Cyt, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-Bcl-2 antibody [100/D5]
    See all Bcl-2 primary antibodies

  • Description

    Mouse monoclonal [100/D5] to Bcl-2

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, Flow Cyt, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Cow, Dog, Chinese hamsterDoes not react with: Rat

  • Immunogen

    Synthetic peptide corresponding to Bcl-2 aa 41-54.
    Sequence:

    GAAPAPGIFSSQPG-Cys


    Database link: P10415
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • IHC: Human tonsil ICC/IF: Human neuroblastoma (SK-N-SH cells) WB: HAP1 cells and HeLa cells Flow Cyt: Jurkat cells

  • General notes

    This product was changed from ascites to tissue culture supernatant on 8th March 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab692 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 (7)
Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.
Flow Cyt
1/10.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 5 µg/ml.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.
Flow Cyt
1/10.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 5 µg/ml.

Target

  • Function

    Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release (PubMed:17418785).

  • Tissue specificity

    Expressed in a variety of tissues.

  • Involvement in disease

    A chromosomal aberration involving BCL2 has been found in chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions.

  • Sequence similarities

    Belongs to the Bcl-2 family.

  • Domain

    BH1 and BH2 domains are required for the interaction with BAX and for anti-apoptotic activity.
    The BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.
    The loop between motifs BH4 and BH3 is required for the interaction with NLRP1.

  • Post-translational
    modifications

    Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A).
    Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.
    Monoubiquitinated by PARK2, leading to increase its stability. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome.

  • Cellular localization

    Mitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Apoptosis regulator Bcl 2 antibody
    • Apoptosis regulator Bcl-2 antibody
    • Apoptosis regulator Bcl2 antibody
    • AW986256 antibody
    • B cell CLL/lymphoma 2 antibody
    • B cell leukemia/lymphoma 2 antibody
    • Bcl-2 antibody
    • Bcl2 antibody
    • BCL2_HUMAN antibody
    • C430015F12Rik antibody
    • D630044D05Rik antibody
    • D830018M01Rik antibody
    • Leukemia/lymphoma, B-cell, 2 antibody
    • Oncogene B-cell leukemia 2 antibody
    • PPP1R50 antibody
    • Protein phosphatase 1 regulatory subunit 50 antibody
    • Protein phosphatase 1, regulatory subunit 50 antibody
    see all

Images

  • Western blot - Anti-Bcl-2 antibody [100/D5] (ab692)
    Western blot - Anti-Bcl-2 antibody [100/D5] (ab692)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: BCL2 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lanes 1 - 3: Merged signal (red and green). Green - ab692 observed at 26 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab692 was shown to specifically react with BCL2 when BCL2 knockout samples were used. Wild-type and BCL2 knockout samples were subjected to SDS-PAGE. Ab692 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 antibody [100/D5] (ab692)
    Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 antibody [100/D5] (ab692)

    ab692 staining Bcl-2 in SK-N-SH cells treated with (R)-(-)-Deprenyl hydrochloride (Selegiline hydrochloride) (ab120604), by ICC/IF. Increase of Bcl-2 expression correlates with increased concentration of (R)-(-)-Deprenyl hydrochloride (Selegiline hydrochloride), as described in literature.
    The cells were incubated at 37°C for 3h in media containing different concentrations of ab120604 ((R)-(-)-Deprenyl hydrochloride (Selegiline hydrochloride)) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab692 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A anti-mouse DyLight 488 polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [100/D5] (ab692)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [100/D5] (ab692)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Bcl-2 with ab692 at 1/100 dilution. Samples were incubated with primary antibody for 30-45 minutes at RT.

  • Flow Cytometry - Anti-Bcl-2 antibody [100/D5] (ab692)
    Flow Cytometry - Anti-Bcl-2 antibody [100/D5] (ab692)

    Overlay histogram showing Jurkat cells stained with ab692 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab692, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References (211)

Publishing research using ab692? Please let us know so that we can cite the reference in this datasheet.

ab692 has been referenced in 211 publications.

  • Ding R  et al. Antrodia camphorata extract (ACE)-induced apoptosis is associated with BMP4 expression and p53-dependent ROS generation in human colon cancer cells. J Ethnopharmacol 268:113570 (2021). PubMed: 33181285
  • Yoon C  et al. PI3K/Akt pathway and Nanog maintain cancer stem cells in sarcomas. Oncogenesis 10:12 (2021). PubMed: 33468992
  • Rong JH  et al. Lobaplatin Enhances Radioactive 125I Seed-Induced Apoptosis and Anti-Proliferative Effect in Non-Small Cell Lung Cancer by Suppressing the AKT/mTOR Pathway. Onco Targets Ther 14:289-300 (2021). PubMed: 33469307
  • Gui C  et al. Total flavone extract from Ampelopsis megalophylla induces apoptosis in the MCF-7 cell line. Int J Oncol 58:409-418 (2021). PubMed: 33469684
  • Liu K & Ma R MicroRNA-615-5p regulates the proliferation and apoptosis of breast cancer cells by targeting HSF1. Exp Ther Med 21:192 (2021). PubMed: 33488801
View all Publications for this product

Customer reviews and Q&As

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1-10 of 16 Abreviews or Q&A

Immunocytochemistry abreview for Anti-Bcl-2 antibody [100/D5]

 Good
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Cardiomyocytes)
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
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 Mar 13 2018

Western blot abreview for Anti-Bcl-2 [100/D5] antibody

 Poor
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Mouse Tissue lysate - whole (Heart)
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°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 Oct 21 2013

Western blot abreview for Anti-Bcl-2 antibody [100/D5]

 Poor
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Rat Tissue lysate - whole (Heart)
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°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 Oct 14 2013

Western blot abreview for Anti-Bcl-2 antibody [100/D5]

 Average
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Human Tissue lysate - whole (Heart)
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°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 Oct 14 2013

Western blot abreview for Anti-Bcl2 antibody [100/D5]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma Initiating Cells (Primary cells))
Loading amount
30 µg
Specification
Glioblastoma Initiating Cells (Primary cells)
Gel Running Conditions
Reduced Denaturing (12.5)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°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 16 2011

Western blot abreview for Anti-Bcl2 antibody [100/D5]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Tissue lysate - whole (human islet lysate)
Loading amount
20 µg
Specification
human islet lysate
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°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.

Mr. Hao Wu

Verified customer

Submitted Nov 10 2011

Western blot abreview for Anti-Bcl2 antibody [100/D5]

 Good
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa Cells)
Loading amount
20 µg
Specification
HeLa Cells
Gel Running Conditions
Non-reduced Denaturing (10)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°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 Aug 23 2011

Question

Inquiry: I'm interested if there is a possibility to get small amount of antibodies for test purposes. If yes, what is the amount, concentration and price for that. We are interested for -Anti-CDKN1A interacting zinc finger protein 1 antibody (ab102013) -Anti-p53 antibody (ab1101) -Anti-Bcl2 antibody [100/D5] (ab692) -Anti-Bax antibody (ab7977) Thank you for answer Regards

Read More
Answer

Thank you for contacting us. Unfortunately, as we carry over 70,000 products, it simply isn't feasible for us to keep small sample sizes of our products.

However, we are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information. More information on this offer can be found from the following link:

https://www.abcam.com/collaboratordiscount

In order to qualify for this offer you need to be issued with a testing discount code prior to the purchase of the antibody to be tested. Please do therefore let us know if you would like to participate in this scheme.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More

Question

Hi,



I have some antibodies from you, and their datasheets say that I need to store them at 4C, for example, ab98711 and ab692.

However, most of antibodies from abcam need to be stored at –20C or –80C.



So, my question is what is the estimated shelf life of the product if I store it at 4C as its datasheet says.

I’m worried whether it is correct that they are loosing their affinity or denatured with a higher rate compared to other antibodies kept at –20C as time goes by.
And, what happens if I freeze it?

Thank you in advance.

Read More
Answer

Thank you for your enquiry.

Estimated shelf life of antibodies stored at 4 degrees C is about 1 year, although frequently they last much longer.

I recommend that you follow the storage conditions as noted on the product datasheet. Freezing an antibody that is intended to only be stored at 4 degrees C may lead to denaturing of the antibody.

I hope this is helpful. Please contact me again if you have any further questions.

Read More

Question

Hi.

I have frozen Ab692 by a mistake. The freezer's temperature is around
-30 degrees. What should I do? Should I take it out and put it in -80 or
transfer it to a refrigerator at 4 degrees? Do you think the antibody is
still usable for IHC?

Regards,

Read More
Answer

Thank you for your enquiry. I am sorry to hear you have some concerns regarding the storage of the antibody.

I would suggest to take the antibody out of the freezer and put it at +4oC. The antibody solution does contain some BSA, which can act as a cryoprotectant, so the antibody may be ok and I would certainly recommend it is worth trying in an experiment. However, regrettably I am sorry we would not longer be able to guarantee how well this will work.

If it has lost some activity, you could try using it at a higher concentration.

I hope this will be helpful. Please do not hesitate to let me know if you have any further questions. Good luck with the experiments.

Read More

1-10 of 16 Abreviews or Q&A

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