Caspase 9 Antibody | Affinity Biosciences

Product:	Caspase 9 Antibody
Catalog:	AF6348
Description:	Rabbit polyclonal antibody to Caspase 9
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Sheep
Mol.Wt.:	46kDa; 46kD(Calculated).
Uniprot:	P55211
RRID:	AB_2835042

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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.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

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

Prediction:

Pig(88%), Bovine(82%), Horse(91%), Sheep(90%)

Clonality:

Polyclonal

Specificity:

Caspase 9 Antibody detects endogenous levels of total Caspase 9.

RRID:

AB_2835042
Cite Format: Affinity Biosciences Cat# AF6348, RRID:AB_2835042.

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

APAF-3; APAF3; Apoptosis related cysteine peptidase; Apoptotic protease Mch-6; Apoptotic protease-activating factor 3; CASP-9; CASP9; CASP9_HUMAN; Caspase 9 apoptosis related cysteine peptidase; Caspase 9 Dominant Negative; Caspase 9c; Caspase-9; Caspase-9 subunit p10; ICE LAP6; ICE like apoptotic protease 6; ICE-LAP6; ICE-like apoptotic protease 6; MCH6; PPP1R56; protein phosphatase 1, regulatory subunit 56; RNCASP9;

Immunogens

Immunogen:

Uniprot:

P55211(CASP9_HUMAN) >>Visit HPA database.

Gene(ID):

CASP9(842)

Expression:

P55211 CASP9_HUMAN:

Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes.

Description:

This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce 2 subunits, large and small, that dimerize to form the active enzyme.

Sequence:

P55211 CASP9_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MDEADRRLLRRCRLRLVEELQVDQLWDALLSRELFRPHMIEDIQRAGSGSRRDQARQLIIDLETRGSQALPLFISCLEDTGQDMLASFLRTNRQAAKLSKPTLENLTPVVLRPEIRKPEVLRPETPRPVDIGSGGFGDVGALESLRGNADLAYILSMEPCGHCLIINNVNFCRESGLRTRTGSNIDCEKLRRRFSSLHFMVEVKGDLTAKKMVLALLELAQQDHGALDCCVVVILSHGCQASHLQFPGAVYGTDGCPVSVEKIVNIFNGTSCPSLGGKPKLFFIQACGGEQKDHGFEVASTSPEDESPGSNPEPDATPFQEGLRTFDQLDAISSLPTPSDIFVSYSTFPGFVSWRDPKSGSWYVETLDDIFEQWAHSEDLQSLLLRVANAVSVKGIYKQMPGCFNFLRKKLFFKTS

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

Sheep

Pig

Bovine

Rabbit

Xenopus

Zebrafish

Dog

Chicken

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

PTMs - P55211 As Substrate

P55211

Site	PTM Type	Enzyme	Source
K97	Ubiquitination		Uniprot
S99	Phosphorylation	P17612 (PRKACA)	Uniprot
K100	Ubiquitination		Uniprot
T107	Phosphorylation		Uniprot
K117	Ubiquitination		Uniprot
T125	Phosphorylation	P27361 (MAPK3) , P28482 (MAPK1) , Q13627 (DYRK1A) , P06493 (CDK1)	Uniprot
S133	Phosphorylation		Uniprot
S144	Phosphorylation	Q05513 (PRKCZ)	Uniprot
Y153	Phosphorylation	P00519 (ABL1) , A0A173G4P4 (Abl fusion)	Uniprot
S175	Phosphorylation		Uniprot
S183	Phosphorylation	P17612 (PRKACA)	Uniprot
K189	Ubiquitination		Uniprot
S195	Phosphorylation	P17612 (PRKACA)	Uniprot
S196	Phosphorylation	Q9Y243 (AKT3) , P31751 (AKT2) , P31749 (AKT1) , Q13237 (PRKG2)	Uniprot
K204	Ubiquitination		Uniprot
T208	Phosphorylation		Uniprot
K210	Ubiquitination		Uniprot
K211	Ubiquitination		Uniprot
Y251	Phosphorylation		Uniprot
K278	Ubiquitination		Uniprot
T301	Phosphorylation		Uniprot
S302	Phosphorylation		Uniprot
S307	Phosphorylation		Uniprot
S310	Phosphorylation		Uniprot
K394	Ubiquitination		Uniprot
Y397	Phosphorylation		Uniprot

Research Backgrounds

P55211_CASP9_HUMAN

Function:

Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Promotes DNA damage-induced apoptosis in a ABL1/c-Abl-dependent manner. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).

Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9.

PTMs:

Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase-3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events.

Phosphorylated at Thr-125 by MAPK1/ERK2. Phosphorylation at Thr-125 is sufficient to block caspase-9 processing and subsequent caspase-3 activation. Phosphorylation on Tyr-153 by ABL1/c-Abl; occurs in the response of cells to DNA damage.

Tissue Specificity:

Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes.

Subunit Structure:

Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 35 kDa (p35) and a 10 kDa (p10) subunit. Caspase-9 and APAF1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome C and ATP. Interacts (inactive form) with EFHD2. Interacts with HAX1. Interacts with BIRC2/c-IAP1, XIAP/BIRC4, BIRC5/survivin, BIRC6/bruce and BIRC7/livin. Interacts with ABL1 (via SH3 domain); the interaction is direct and increases in the response of cells to genotoxic stress and ABL1/c-Abl activation. Interacts with NleF from pathogenic E.coli.

Family&Domains:

Belongs to the peptidase C14A family.

Research Fields

· Cellular Processes > Cell growth and death > p53 signaling pathway. (View pathway)

· 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 > PI3K-Akt signaling pathway. (View pathway)

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

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· Human Diseases > Neurodegenerative diseases > Parkinson's disease.

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

· Human Diseases > Neurodegenerative diseases > Huntington's disease.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

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

· Human Diseases > Infectious diseases: Viral > Influenza A.

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

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

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

· Human Diseases > Cancers: Specific types > Endometrial 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 > Non-small cell lung cancer. (View pathway)

· Human Diseases > Cardiovascular diseases > Viral myocarditis.

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

References

1). 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.

2). Formation of di-cysteine acrolein adduct decreases cytotoxicity of acrolein by ROS alleviation and apoptosis intervention. Journal of Hazardous Materials, 2020 (PubMed: 31780296) [IF=13.6]

Application: WB Species: Human Sample: HBE (A) and Caco-2 (B) cells

Fig. 8 Effect of acrolein (ACR) or its adduct ACR-di-Cys with/without addition of 1mM cysteine on the cleavage of caspase-3, caspase-8 and PARP in HBE (A) and Caco-2 (B) cells. Left: western blot. Right: The data were expressed as relative intensity to GAPDH; Error bars represent standard deviation (n=3); Different letters indicate significant differences (p < 0.05) between treatments. 42 J

Application: WB Species: Human Sample: HBE and Caco-2 cells

Fig. 9 Effect of acrolein (ACR) or its adduct ACR-di-Cys with/without addition of 1mM cysteine on the expression of Bax, Bcl-2, cytochrome c and the cleavage of caspase-9 in HBE (A) and Caco-2 (B) cells. Left: western blot. Right: The data were expressed as relative intensity to GAPDH; Error bars represent standard deviation (n=3); Different letters indicate significant differences (p < 0.05) between treatments.

3). Echinatin inhibits the growth and metastasis of human osteosarcoma cells through Wnt/β-catenin and p38 signaling pathways. Pharmacological Research, 2023 (PubMed: 37023991) [IF=9.3]

Application: WB Species: Human Sample: OS cells

Fig. 4. Ecn inhibits the migration and invasion of OS cells. (A) The effect of Ecn on the migration of OS cells (Wound healing assay, 100 ×). (B) The effect of Ecn on the migration of OS cells (Transwell assay, 100 ×). (C) The effect of Ecn on the invasion of OS cells (Matrigel-coated Transwell assay, 100 ×). (D) The effect of Ecn on the protein level of MMP2, MMP7, MMP9, Snail, Vimentin, N-Cadherin and E-Cadherin in OS cells (Western blot). ##P

4). Increased autophagy in EOC re-ascites cells can inhibit cell death and promote drug resistance. Cell Death & Disease, 2018 (PubMed: 29549251) [IF=9.0]

Application: IHC Species: human Sample: EOC ascites cells

Fig. 2| Autophagy is increased and apoptosis is reduced in the EOC re-ascites group. a IHC analysis of LC-3, Belin-1, CASP-9, and c-CASP-3 expression in EOC ascites cells (scale bar =30µm). b, c Western blot analysis of autophagy/apoptosis-related proteins expressed in the chemosensitive group and re-ascites group (***p<0.001, **p<0.01, *p<0.05). d Immunofluorescence analysis of the chemosensitive group and re-ascites group with LC3 (green) and LAMP2 (red) staining. Nuclei were counterstained with DAPI (blue). e qRT-PCR analysis of the average relative mRNA expression levels of autophagy/apoptosis-related genes in the chemosensitive group and re-ascites group (**p<0.01, *p<0.05)

Application: IF/ICC Species: human Sample: ascites cells

Fig. 5 |Apoptosis is increased in the EOC chemosensitive group. a, b IHC and IF analysis of apoptotic protein expression in ovarian cancer samples. c, d Western blot analysis of apoptotic proteins expressed in the no-chemotherapy group and chemosensitive group (***p<0.001,**p<0.01, *p<0.05). e qRT-PCR analysis of the average relative mRNA expression levels of apoptosis-related genes in the no-chemotherapy group and chemosensitive group (*p<0.05)

Application: WB Species: human Sample: ascites cells

5). Cytotoxicity of adducts formed between quercetin and methylglyoxal in PC-12 cells. Food Chemistry, 2021 (PubMed: 33706136) [IF=8.8]

Application: WB Species: Rat Sample: PC-12 cells

Fig. 5. Effect of treatments of MGO, Que-mono-MGO, and Que-di-MGO on the expression levels of apoptotic markers and components of AKT and Nrf2-HO-1/NQO-1 signaling pathways. Significant differences (p < 0.05) between samples of different treatments are marked with different letters on each column.

6). LNC473 regulating APAF1 IRES-dependent translation via competitive sponging miR574 and miR15b: Implications in colorectal cancer. Molecular Therapy-Nucleic Acids, 2020 (PubMed: 32784109) [IF=8.8]

Application: WB Species: Human Sample: HCT116 and SW480 cells

Figure 6. LNC473-miR574/miR15b-APAF1 Signaling Axis in HCT116 and SW480 Cells (A) IF and ISH combination assay revealing the co-localization of APAF1 protein with included ncRNAs in HCT116 cells. Scale bars, 5 mm. (B and C) The levels of APAF1 mRNA (B) and protein (C) were determined after interfering LNC473 expression in HCT116 and SW480 cells by qPCR and IF assays. Scale bars, 20 mm. (D) The expression of apoptosis-related proteins including APAF1 was detected after interfering LNC473 expression in HCT116 and SW480 cells by western blot assay. (E and F) Rescue experiments showing the APAF1 levels in HCT116 and SW480 cells with exposure to the co-transfection of pcDH-LNC473 vector and miR574-5p or miR15b-5p mimic by qPCR (E) and western blot (F) assays. (G) Pattern diagram of APAF1-CDS and APAF1-IRES-CDS vectors. (H and I) APAF1 protein expression was determined in HCT116 and SW480 cells treated with APAF1-IRES-CDS vector, or pcDH-LNC473 and APAF1-IRES-CD co-transfection by western blot (H) and IF assays (I). Scale bars, 5 mm. (J) The percentage (%) of cell apoptosis in cells upon co-overexpressing APAF1-CDS or APAF1-IRE-CDS and LNC473 as assayed by flow cytometry. All tests were performed at least three times. Data were expressed as mean ± SD. ns (nonsignificant), p > 0.05; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

7). Long noncoding RNAs regulated spermatogenesis in varicocele‐induced spermatogenic dysfunction. CELL PROLIFERATION, 2022 (PubMed: 35297519) [IF=8.5]

Application: WB Species: Human Sample: spermatogenic cell

FIGURE 13Validation of regulated signalling pathways, spermatogenic cell apoptosis and proliferation, and meiotic spermatocytes by Western blot. Representative Western blot images of PI3K, Akt, p-Akt, caspase-9, Bcl-2, Bax, PCNA, PLZF, REC8, STRA8, and SYCP3. (B) Statistical analysis of band intensity by Student's t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. NS, not significant; Sham, sham group; Surgical treatment, surgical treatment group; VC, varicocele group

8). Synthesis, characterization, and cytotoxicity analysis of selenium nanoparticles stabilized by Morchella sextelata polysaccharide. International Journal of Biological Macromolecules, 2023 (PubMed: 37247714) [IF=8.2]

9). Development of anisamide-targeted PEGylated gold nanorods to deliver epirubicin for chemo-photothermal therapy in tumor-bearing mice. International Journal of Nanomedicine, 2019 (PubMed: 30880982) [IF=8.0]

Application: WB Species: Sample: PC-3 cells

Figure 6 |The expression of caspase 3, cleaved caspase 3 (phosphorylated form), caspase 9, cleaved caspase 9 (phosphorylated form), Bcl-2, Bax, and β-actin was determined using Western blotting.

10). Quercetin induces autophagy via FOXO1-dependent pathways and autophagy suppression enhances quercetin-induced apoptosis in PASMCs in hypoxia. FREE RADICAL BIOLOGY AND MEDICINE, 2017 (PubMed: 27979659) [IF=7.4]

Application: WB Species: rat Sample:

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Caspase 9 Antibody - #AF6348