Bim Antibody | Affinity Biosciences

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AF0121 at 1/100 staining human Colorectal cancer tissue by IHC-P.

$Figure 5 DHW-221-triggered mitochondrial apoptosis is linked to Akt-mediated FOXO3a nuclear translocation in A549/Taxol cells. (A, B) Comparison and the expression levels of PI3K/Akt/FOXO3a signaling pathway related proteins in A549 and A549/Taxol cells were determined by Western blot, in the presence and absence of DHW-221 and GDC-0980. Statistical comparisons were performed with unpaired Student’s t-test (n = 3) in Panel (A). *p < 0.05, **p < 0.01 versus A549. Statistical comparisons were performed with one-way ANOVA followed by Dunnett’s post-hoc test for multiple comparisons in Panel (B) (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001 versus control. (C, E) The FOXO3a and p-FOXO3a expressions in the nuclear and cytoplasmic fractions of A549/Taxol cells were detected by Western blot. Proliferating cell nuclear antigen (PCNA) was selected as nucleoprotein internal control. The quantitative results were shown in Panel (E). (D) Immunofluorescence staining of FOXO3a in A549/Taxol cells was carried out to evaluate the effect of DHW-221 on FOXO3a nuclear translocation. Scale bar = 20 µm. The histograms indicated the percentage of the cells in each condition exhibiting FOXO3a nuclear mean fluorescence intensity (positive cells, green fluorescence) by ImageJ. (F) FOXO3a degradation in A549/Taxol cells with or without DHW-221 co-treatment in different time spot when protein biosynthesis was blocked with 20 µM cycloheximide (CHX). FOXO3a stability was analyzed relative to control by ImageJ software. (G) FOXO3a proteins levels in the presence of MG132 (0.20 μM) or 2.40 μM DHW-221-treated A549/Taxol cells at 24 h. Statistical comparisons were performed with one-way ANOVA followed by Dunnett’s post-hoc test for multiple comparisons (n = 3). Data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 versus control.$

Figure 5 DHW-221-triggered mitochondrial apoptosis is linked to Akt-mediated FOXO3a nuclear translocation in A549/Taxol cells.

Product:	Bim Antibody
Catalog:	AF0121
Description:	Rabbit polyclonal antibody to Bim
Application:	WB IHC
Reactivity:	Human, Mouse, Rat, Monkey
Prediction:	Pig, Horse, Sheep, Rabbit, Dog
Mol.Wt.:	25kDa,22kDa; 22kD(Calculated).
Uniprot:	O43521
RRID:	AB_2833305

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:3000, IHC 1:50-1:200
*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,Monkey

Prediction:

Pig(100%), Horse(86%), Sheep(100%), Rabbit(100%), Dog(100%)

Clonality:

Polyclonal

Specificity:

Bim Antibody detects endogenous levels of total Bim.

RRID:

AB_2833305
Cite Format: Affinity Biosciences Cat# AF0121, RRID:AB_2833305.

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

BCL2 like 11; B2L11_HUMAN; BAM; Bcl 2 interacting protein Bim; Bcl 2 related ovarian death agonist; Bcl-2-like protein 11; BCL2 interacting mediator of cell death; BCL2 like 11 (apoptosis facilitator); BCL2 like protein 11; Bcl2-interacting mediator of cell death; Bcl2-L-11; Bcl2l11; BIM alpha6; BIM; BIM beta6; BIM beta7; BimEL; BimL; BOD;

Immunogens

Immunogen:

Uniprot:

O43521(B2L11_HUMAN) >>Visit HPA database.

Gene(ID):

BCL2L11(10018)

Expression:

O43521 B2L11_HUMAN:

Isoform BimEL, isoform BimL and isoform BimS are the predominant isoforms and are widely expressed with tissue-specific variation. Isoform Bim-gamma is most abundantly expressed in small intestine and colon, and in lower levels in spleen, prostate, testis, heart, liver and kidney.

Description:

Bim a pro-apoptotic member of the BCL-2 protein family. Interacts with other members of the BCL-2 protein family, including BCL2, BCL2L1/BCL-X(L), and MCL1, and act as an apoptotic activator. Its expression can be induced by nerve growth factor (NGF), as well as by the forkhead transcription factor FKHR-L1, which suggests a role in neuronal and lymphocyte apoptosis. Transgenic studies in the mouse suggested that this protein functions as an essential initiator of the apoptosis in thymocyte-negative selection. Eight alternatively spliced transcript variants of this gene have been reported.

Sequence:

O43521 B2L11_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MAKQPSDVSSECDREGRQLQPAERPPQLRPGAPTSLQTEPQGNPEGNHGGEGDSCPHGSPQGPLAPPASPGPFATRSPLFIFMRRSSLLSRSSSGYFSFDTDRSPAPMSCDKSTQTPSPPCQAFNHYLSAMASMRQAEPADMRPEIWIAQELRRIGDEFNAYYARRVFLNNYQAAEDHPRMVILRLLRYIVRLVWRMH

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

Pig

100

Sheep

100

Dog

100

Rabbit

100

Horse

Bovine

Xenopus

Zebrafish

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O43521 As Substrate

O43521

Site	PTM Type	Enzyme	Source
	Phosphorylation		Uniprot
C12	S-Nitrosylation		Uniprot
S59	Phosphorylation		Uniprot
S65	Phosphorylation		Uniprot
S69	Phosphorylation	Q16539 (MAPK14) , P45983 (MAPK8) , P45984 (MAPK9) , P53779 (MAPK10) , P28482 (MAPK1) , P27361 (MAPK3)	Uniprot
S77	Phosphorylation		Uniprot
S87	Phosphorylation	P31749 (AKT1)	Uniprot
S93	Phosphorylation	O14965 (AURKA)	Uniprot
S94	Phosphorylation	O14965 (AURKA)	Uniprot
Y96	Phosphorylation		Uniprot
S98	Phosphorylation	O14965 (AURKA)	Uniprot
S104	Phosphorylation	P45983 (MAPK8)	Uniprot
S113	Phosphorylation		Uniprot
T116	Phosphorylation	P53779 (MAPK10) , P45983 (MAPK8)	Uniprot
S118	Phosphorylation	P45983 (MAPK8)	Uniprot

Research Backgrounds

O43521_B2L11_HUMAN

Function:

Induces apoptosis and anoikis. Isoform BimL is more potent than isoform BimEL. Isoform Bim-alpha1, isoform Bim-alpha2 and isoform Bim-alpha3 induce apoptosis, although less potent than isoform BimEL, isoform BimL and isoform BimS. Isoform Bim-gamma induces apoptosis. Isoform Bim-alpha3 induces apoptosis possibly through a caspase-mediated pathway. Isoform BimAC and isoform BimABC lack the ability to induce apoptosis.

PTMs:

Phosphorylation at Ser-69 by MAPK1/MAPK3 leads to interaction with TRIM2 and polyubiquitination, followed by proteasomal degradation. Deubiquitination catalyzed by USP27X stabilizes the protein (By similarity).

Ubiquitination by TRIM2 following phosphorylation by MAPK1/MAPK3 leads to proteasomal degradation. Conversely, deubiquitination catalyzed by USP27X stabilizes the protein.

Subcellular Location:

Endomembrane system>Peripheral membrane protein.
Note: Associated with intracytoplasmic membranes.

Mitochondrion.
Note: Translocates from microtubules to mitochondria on loss of cell adherence.

Mitochondrion.

Mitochondrion.

Mitochondrion.

Tissue Specificity:

Subunit Structure:

Forms heterodimers with a number of antiapoptotic Bcl-2 proteins, including MCL1, BCL2, BCL2L1 isoform Bcl-X(L), BCL2A1/BFL-1, BHRF1, and BCL2L2/BCLW. Isoform BimS and isoform Bim-alpha3 interact with BAX; this interaction may lead to BAX activation through conformational change. Does not heterodimerize with proapoptotic proteins such as BAD, BOK or BAK. Identified in a complex containing BCL2L11, DYNLL1 and BCL2L1 isoform Bcl-X(L); BH3 integrity is required for BCL2L1-binding. Interacts with YWHAZ. When phosphorylated, interacts with TRIM2; this interaction is associated with ubiquitination and degradation. Interacts with MCL1; may sequester BCL2L11 to prevent its pro-apoptotic activity. When phosphorylated, isoform BimEL interacts with USP27X; this interaction leads to BCL2L11 deubiquitination and stabilization. Interacts with GIMAP5.

Family&Domains:

The BH3 motif is required for interaction with Bcl-2 proteins and cytotoxicity.

Belongs to the Bcl-2 family.

Research Fields

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

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

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

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

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

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· 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)

References

1). DHW-221, a Dual PI3K/mTOR Inhibitor, Overcomes Multidrug Resistance by Targeting P-Glycoprotein (P-gp/ABCB1) and Akt-Mediated FOXO3a Nuclear Translocation in Non-small Cell Lung Cancer. Frontiers in Oncology, 2022 (PubMed: 35646704) [IF=4.7]

Application: WB Species: Human Sample: A549 cells

Figure 5 DHW-221-triggered mitochondrial apoptosis is linked to Akt-mediated FOXO3a nuclear translocation in A549/Taxol cells. (A, B) Comparison and the expression levels of PI3K/Akt/FOXO3a signaling pathway related proteins in A549 and A549/Taxol cells were determined by Western blot, in the presence and absence of DHW-221 and GDC-0980. Statistical comparisons were performed with unpaired Student’s t-test (n = 3) in Panel (A). *p < 0.05, **p < 0.01 versus A549. Statistical comparisons were performed with one-way ANOVA followed by Dunnett’s post-hoc test for multiple comparisons in Panel (B) (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001 versus control. (C, E) The FOXO3a and p-FOXO3a expressions in the nuclear and cytoplasmic fractions of A549/Taxol cells were detected by Western blot. Proliferating cell nuclear antigen (PCNA) was selected as nucleoprotein internal control. The quantitative results were shown in Panel (E). (D) Immunofluorescence staining of FOXO3a in A549/Taxol cells was carried out to evaluate the effect of DHW-221 on FOXO3a nuclear translocation. Scale bar = 20 µm. The histograms indicated the percentage of the cells in each condition exhibiting FOXO3a nuclear mean fluorescence intensity (positive cells, green fluorescence) by ImageJ. (F) FOXO3a degradation in A549/Taxol cells with or without DHW-221 co-treatment in different time spot when protein biosynthesis was blocked with 20 µM cycloheximide (CHX). FOXO3a stability was analyzed relative to control by ImageJ software. (G) FOXO3a proteins levels in the presence of MG132 (0.20 μM) or 2.40 μM DHW-221-treated A549/Taxol cells at 24 h. Statistical comparisons were performed with one-way ANOVA followed by Dunnett’s post-hoc test for multiple comparisons (n = 3). Data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 versus control.

2). miR-17-5p attenuates kidney ischemia-reperfusion injury by inhibiting the PTEN and BIM pathways. Annals of Translational Medicine, 2021 (PubMed: 34790751)

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Bim Antibody - #AF0121