Product: Bcl-2 Antibody
Catalog: AF6139
Description: Rabbit polyclonal antibody to Bcl-2
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat, Chinese Mitten Crab
Prediction: Horse, Dog
Mol.Wt.: 26kDa; 26kD(Calculated).
Uniprot: P10415
RRID: AB_2835021

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat,Chinese Mitten Crab
Prediction:
Horse(100%), Dog(86%)
Clonality:
Polyclonal
Specificity:
Bcl-2 Antibody detects endogenous levels of total Bcl-2.
RRID:
AB_2835021
Cite Format: Affinity Biosciences Cat# AF6139, RRID:AB_2835021.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

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

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P10415 BCL2_HUMAN:

Expressed in a variety of tissues.

Description:
This gene encodes an integral outer mitochondrial membrane protein that blocks the apoptotic death of some cells such as lymphocytes. Constitutive expression of BCL2, such as in the case of translocation of BCL2 to Ig heavy chain locus, is thought to be the cause of follicular lymphoma. Two transcript variants, produced by alternate splicing, differ in their C-terminal ends.
Sequence:
MAHAGRTGYDNREIVMKYIHYKLSQRGYEWDAGDVGAAPPGAAPAPGIFSSQPGHTPHPAASRDPVARTSPLQTPAAPGAAAGPALSPVPPVVHLTLRQAGDDFSRRYRRDFAEMSSQLHLTPFTARGRFATVVEELFRDGVNWGRIVAFFEFGGVMCVESVNREMSPLVDNIALWMTEYLNRHLHTWIQDNGGWDAFVELYGPSMRPLFDFSWLSLKTLLSLALVGACITLGAYLGHK

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Horse
100
Dog
86
Pig
0
Bovine
0
Sheep
0
Xenopus
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P10415 As Substrate

Site PTM Type Enzyme
Y9 Phosphorylation
K22 Ubiquitination
S24 Phosphorylation
T56 Phosphorylation Q16539 (MAPK14) , P06493 (CDK1) , P53779 (MAPK10) , P28482 (MAPK1) , P27361 (MAPK3)
T69 Phosphorylation P45983 (MAPK8)
S70 Phosphorylation P27361 (MAPK3) , P06493 (CDK1) , P53779 (MAPK10) , P17252 (PRKCA) , Q00534 (CDK6) , P28482 (MAPK1) , P45983 (MAPK8)
T74 Phosphorylation P28482 (MAPK1) , P53779 (MAPK10) , P27361 (MAPK3)
S87 Phosphorylation Q16539 (MAPK14) , P45983 (MAPK8) , Q00534 (CDK6) , P27361 (MAPK3) , P28482 (MAPK1) , P53779 (MAPK10)
C158 S-Nitrosylation
C229 S-Nitrosylation
Y235 Phosphorylation

Research Backgrounds

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.

PTMs:

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) (By similarity).

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 PRKN, leading to increase its stability. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome.

Subcellular Location:

Mitochondrion outer membrane>Single-pass membrane protein. Nucleus membrane>Single-pass membrane protein. Endoplasmic reticulum membrane>Single-pass membrane protein.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in a variety of tissues.

Subunit Structure:

Forms homodimers, and heterodimers with BAX, BAD, BAK and Bcl-X(L). Heterodimerization with BAX requires intact BH1 and BH2 motifs, and is necessary for anti-apoptotic activity. Interacts with EI24 (By similarity). Also interacts with APAF1, BBC3, BCL2L1, BNIPL, MRPL41 and TP53BP2. Binding to FKBP8 seems to target BCL2 to the mitochondria and probably interferes with the binding of BCL2 to its targets. Interacts with BAG1 in an ATP-dependent manner. Interacts with RAF1 (the 'Ser-338' and 'Ser-339' phosphorylated form). Interacts (via the BH4 domain) with EGLN3; the interaction prevents the formation of the BAX-BCL2 complex and inhibits the anti-apoptotic activity of BCL2. Interacts with G0S2; this interaction also prevents the formation of the anti-apoptotic BAX-BCL2 complex. Interacts with RTL10/BOP. Interacts with the SCF(FBXO10) complex. Interacts (via the loop between motifs BH4 and BH3) with NLRP1 (via LRR repeats), but not with NLRP2, NLRP3, NLRP4, PYCARD, nor MEFV. Interacts with GIMAP3/IAN4, GIMAP4/IAN1 and GIMAP5/IAN5 (By similarity).

Family&Domains:

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.

Belongs to the Bcl-2 family.

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis - multiple species.   (View pathway)

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Hedgehog signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.   (View pathway)

· Genetic Information Processing > Folding, sorting and degradation > Protein processing in endoplasmic reticulum.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

· Organismal Systems > Circulatory system > Adrenergic signaling in cardiomyocytes.   (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

· Organismal Systems > Nervous system > Cholinergic synapse.

· Organismal Systems > Endocrine system > Estrogen signaling pathway.   (View pathway)

References

1). Efficient Therapy of Inflammatory Bowel Disease (IBD) with Highly Specific and Durable Targeted Ta2C Modified with Chondroitin Sulfate (TACS). Advanced Materials, 2023 (PubMed: 37224059) [IF=29.4]

Application: WB    Species: Mouse    Sample: colon tissue

Figure 6.TACS prevents apoptosis. C) Western blot analysis of caspase 3, BAX, and Bcl-2 proteins expression in colon tissue homogenate fromdifferent groups.

2). Smooth muscle NF90 deficiency ameliorates diabetic atherosclerotic calcification in male mice via FBXW7-AGER1-AGEs axis. Nature communications, 2024 (PubMed: 38862515) [IF=16.6]

Application: WB    Species: Mouse    Sample:

Fig. 3 VSMC NF90/110 promotes AGEs-mediated VSMC phenotypic switch, apoptosis, and MVs release. a The HAVSMCs were pre-infected with NF90/110 shRNA for NF90/110 silencing and then incubated in osteogenic medium with AGE-BSA (200 µg/ml) for 24 h. Representative Western blot bands of OPN, BMP2, α-SMA and Calponin expressions and quantification (n = 5 per group). BSA: bovine serum albumin. b Immunofluorescence double staining of OPN (red) and α-SMA (green) in atherosclerotic lesion of ApoE−/− NFflox/flox and ApoE−/−NFSM-KO mice with or without diabetes mellitus (DM) and quantification. Blue DAPI shows positions of nuclei (n = 10 per group, Scale bar: 50 μm). c–e The HAVSMCs were pre-infected with NF90/110 shRNA for NF90/110 silencing and then incubated in osteogenic medium with AGEs (200 µg/ml) for 24 h (n = 5 per group). c VSMCs apoptosis were evaluated by TUNEL staining. Red fluorescence indicates apoptotic cells and blue DAPI shows positions of nuclei (Scale bar: 20 μm) and their quantitative analysis. d The apoptotic rate was evaluated by PE/Annexin V double staining and their quantification. e Representative immunoblots of VSMCs with anti- Bax, Bcl-2 and cleaved-caspase 3 antibodies were shown and quantitative analysis. f TUNEL staining in atherosclerotic lesion of ApoE-/-NFflox/flox and ApoE−/−NFSM-KO mice with or without DM and quantification. Red fluorescence represents apoptotic cells and blue DAPI shows positions of nuclei (n = 10 per group, Scale bar: 50 μm). g MVs were isolated from supernatant of cultured medium and measured by protein content (n = 5 per group). Data were presented as mean ± SEM. One-way ANOVA followed by Tukey’s post-test analysis for (a–g). p-values were adjusted for comparisons of multiple means. Source data are provided as a Source Data file.

3). Gut microbiome mediates the protective effects of exercise after myocardial infarction. Microbiome, 2022 (PubMed: 35637497) [IF=15.5]

4). Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. NUCLEIC ACIDS RESEARCH, 2020 (PubMed: 32710621) [IF=14.9]

Application: WB    Species: human    Sample: HepG2

Figure 4. Effect of A22 on anti-apoptosis in 0.5 mM palmitic acid oil (PA) induced cell model. (A) Effect of A22 on cell viability for anti-apoptotic protective effect. (B) Effect of A22 on transcription of BCL-2 and BAX with measurement of mRNA levels. (C) Effect of A22 on protein expressions related with apoptosis (left), which were quantitatively analyzed (right). All the experiments were repeated for three times.

Application: IHC    Species: mouse    Sample: liver

Figure 6. Effect of A22 on alleviating morphological changes of mice livers. (A) Representative images of livers (× 100 magnification) with oil red O staining, H&E staining, Sirius Red staining, and masson staining. (B–D) Ballooning (blue arrow indicated), hepatic steatosis (black arrow indicated), and fibrosis (blue arrow indicated) scores were obtained according to the NAFLD Activity Score (NAS) System as described in Methods. (E) Relative lipid droplet contents were determined. (F) Expressions of BCL-2 in livers of each group were determined by using immunohistochemistry.

5). Mitochondrial metabolism blockade nanoadjuvant reversed immune-resistance microenvironment to sensitize albumin-bound paclitaxel-based chemo-immunotherapy. Acta pharmaceutica Sinica. B, 2024 (PubMed: 39309498) [IF=14.5]

6). Cyanidin-3-O-glucoside restores spermatogenic dysfunction in cadmium-exposed pubertal mice via histone ubiquitination and mitigating oxidative damage. JOURNAL OF HAZARDOUS MATERIALS, 2020 (PubMed: 31796358) [IF=13.6]

Application: WB    Species: Mice    Sample: testis

Figure 9 The representative photographs and grayscale analysis of proteins in testis from mice treated for 30 days involved in MAPK signaling pathway. (A) p53 and p-p53. (B) Bax, Bcl-2, and Bad. (C) Total Caspase 3 and Cleaved-Caspase 3. (D) Full PARP and Cleaved-PARP. Comparison between all groups was evaluated through One-way ANOVA. Mean±SD. n=4. **p<0.01, *p<0.05, compared with control group. ##p<0.01, #p<0.05, compared with Cd group. ns, not significant.

7). Reproductive toxicity of polystyrene microplastics: In vivo experimental study on testicular toxicity in mice. JOURNAL OF HAZARDOUS MATERIALS, 2021 (PubMed: 33087287) [IF=13.6]

Application: WB    Species: mice    Sample: testis tissue

Fig. 9. Bax and Bcl2 protein expression in the testis tissue mice by western blotting. The statistical chart shows the rate of Bax to Bcl2, which takes into account the balance of apoptosis and anti- apoptosis in testis. *P < 0.05, compared with the corresponding control, , $P < 0.05 compared with the M-Dose group.

8). The anti-microbial peptide LL-37/CRAMP levels are associated with acute heart failure and can attenuate cardiac dysfunction in multiple preclinical models of heart failure. Theranostics, 2020 (PubMed: 32483446) [IF=12.4]

Application: WB    Species: Mice    Sample: heart tissue

Figure 3 CRAMP attenuates TAC-induced heart failure. In TAC mice, mCRAMP peptide improved ejection fractions (EF) and fractional shortening (FS) (A, n=7 in Vehicle+Sham, 10 in CRAMP+Sham, 11 in Vehicle+TAC, and 9 in CRAMP+TAC), decreased cell size (B, n=7 in Vehicle+Sham, 10 in CRAMP+Sham, 11 in Vehicle+TAC, and 9 in CRAMP+TAC)(C, n=4 in Vehicle+Sham, 4 in CRAMP+Sham, 5 in Vehicle+TAC, and 5 in CRAMP+TAC), attenuated cardiac fibrosis (D, n=4 in Vehicle+Sham, 4 in CRAMP+Sham, 5 in Vehicle+TAC, and 5 in CRAMP+TAC), decreased collagen 1, the ratios of Bax/BCl2 and cleaved caspase 3/caspase 3 (E, n=3 per group), and reduced ANP and BNP level (F, n=6 per group). *, P<0.05; **, P<0.01; ***, P<0.001.

9). Inhibition of HIPK2 protects stress-induced pathological cardiac remodeling. EBioMedicine, 2022 (PubMed: 36182775) [IF=11.1]

10). Bacillus spore-based oral carriers loading curcumin for the therapy of colon cancer. JOURNAL OF CONTROLLED RELEASE, 2018 (PubMed: 29274436) [IF=10.8]

Application: WB    Species: human    Sample: HT-29 cells

Figure.5 | Apoptosis detection of HT-29 colon cancer cells. (A) Apoptosis detection of HT-29 cells in different groups by flow cytometry, SFM without drug as control; (B) Apoptosis rates of HT-29 cells in different groups. (mean value ± SD, n=3, **p < 0.01, ***p < 0.001, compared with the control group); (C) Western blotting of the Bcl-2, p53, cleaved caspase-9, cleaved caspase-8,cleaved caspase-3; (D) Relative amount of these apoptosis-related proteins in different groups(mean value ± SD, n=3, *p < 0.05, **p < 0.01, ***p < 0.001, compared with the control group).

Application: IF/ICC    Species:    Sample: tumor

Figure.7 | Analysis of apoptosis-related proteins and curcumin plasma concentration after oral administration. (A) Immunofluorescent images and (B) Quantitative expression analysis of apoptosis-related proteins including Bcl-2, cleaved caspase-3 and p53 in control and SPORE-CUR-FA groups (mean value ± SD, n=5 * p < 0.05, ** p < 0.01, *** p < 0.001, compared with control group). (C) Mean curcumin plasma concentration-time profiles in rats after oral administration of CUR and SPORE-CUR-FA at a dose of 80 mg/kg (mean value ± SD, n= 3). (D)Western blotting of Bcl-2, cleaved caspase-3 and p53 in different groups, physiological saline as control; (E) Relative expression amount of these apoptosis-related proteins in different groups(mean value ± SD, n=3, **p < 0.01, ***p < 0.001, compared with control group).

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