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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [12]
- Comments [0]
- Validations
- Western blot [4]
- Other assay [8]
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- Product number
- PA1-015 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- STUB1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- PA1-015 detects human and mouse carboxyl terminus of hsc70-interacting protein (CHIP). PA1-015 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~35 kDa protein representing CHIP from COS-1 cells overexpressing the human gene and a non-specific band at ~85 kDa. PA1-015 was used successfully in the western blot analysis of CHIP using human MCF7 cell lysates. The PA1-015 immunogen is a synthetic peptide corresponding to residues V(218) D E K R K K R D I P D Y L C(232) of human CHIP. This peptide (Cat. # PEP-179) is available for use in neutralization and control experiments.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references Proteome Instability Is a Therapeutic Vulnerability in Mismatch Repair-Deficient Cancer.
On the Mechanism of Hyperthermia-Induced BRCA2 Protein Degradation.
S-Nitrosylation of CHIP Enhances F508Del-CFTR Maturation.
Modulation of heme/substrate binding cleft of neuronal nitric-oxide synthase (nNOS) regulates binding of Hsp90 and Hsp70 proteins and nNOS ubiquitination.
Brain distribution of carboxy terminus of Hsc70-interacting protein (CHIP) and its nuclear translocation in cultured cortical neurons following heat stress or oxygen-glucose deprivation.
Inhibition of hsp70 by methylene blue affects signaling protein function and ubiquitination and modulates polyglutamine protein degradation.
A critical role for CHIP in the aggresome pathway.
Regulation of death-associated protein kinase. Stabilization by HSP90 heterocomplexes.
CHIP (carboxyl terminus of Hsc70-interacting protein) promotes basal and geldanamycin-induced degradation of estrogen receptor-alpha.
CHIP-Hsc70 complex ubiquitinates phosphorylated tau and enhances cell survival.
Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions.
Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions.
McGrail DJ, Garnett J, Yin J, Dai H, Shih DJH, Lam TNA, Li Y, Sun C, Li Y, Schmandt R, Wu JY, Hu L, Liang Y, Peng G, Jonasch E, Menter D, Yates MS, Kopetz S, Lu KH, Broaddus R, Mills GB, Sahni N, Lin SY
Cancer cell 2020 Mar 16;37(3):371-386.e12
Cancer cell 2020 Mar 16;37(3):371-386.e12
On the Mechanism of Hyperthermia-Induced BRCA2 Protein Degradation.
van den Tempel N, Zelensky AN, Odijk H, Laffeber C, Schmidt CK, Brandsma I, Demmers J, Krawczyk PM, Kanaar R
Cancers 2019 Jan 15;11(1)
Cancers 2019 Jan 15;11(1)
S-Nitrosylation of CHIP Enhances F508Del-CFTR Maturation.
Zaman K, Knight J, Hussain F, Cao R, Estabrooks SK, Altawallbeh G, Holloway K, Jafri A, Sawczak V, Li Y, Getsy P, Sun F, Raffay T, Cotton C, Brodsky JL, Periasamy A, Lewis SJ, Gaston B
American journal of respiratory cell and molecular biology 2019 Dec;61(6):765-775
American journal of respiratory cell and molecular biology 2019 Dec;61(6):765-775
Modulation of heme/substrate binding cleft of neuronal nitric-oxide synthase (nNOS) regulates binding of Hsp90 and Hsp70 proteins and nNOS ubiquitination.
Peng HM, Morishima Y, Pratt WB, Osawa Y
The Journal of biological chemistry 2012 Jan 6;287(2):1556-65
The Journal of biological chemistry 2012 Jan 6;287(2):1556-65
Brain distribution of carboxy terminus of Hsc70-interacting protein (CHIP) and its nuclear translocation in cultured cortical neurons following heat stress or oxygen-glucose deprivation.
Anderson LG, Meeker RB, Poulton WE, Huang DY
Cell stress & chaperones 2010 Sep;15(5):487-95
Cell stress & chaperones 2010 Sep;15(5):487-95
Inhibition of hsp70 by methylene blue affects signaling protein function and ubiquitination and modulates polyglutamine protein degradation.
Wang AM, Morishima Y, Clapp KM, Peng HM, Pratt WB, Gestwicki JE, Osawa Y, Lieberman AP
The Journal of biological chemistry 2010 May 21;285(21):15714-23
The Journal of biological chemistry 2010 May 21;285(21):15714-23
A critical role for CHIP in the aggresome pathway.
Sha Y, Pandit L, Zeng S, Eissa NT
Molecular and cellular biology 2009 Jan;29(1):116-28
Molecular and cellular biology 2009 Jan;29(1):116-28
Regulation of death-associated protein kinase. Stabilization by HSP90 heterocomplexes.
Zhang L, Nephew KP, Gallagher PJ
The Journal of biological chemistry 2007 Apr 20;282(16):11795-804
The Journal of biological chemistry 2007 Apr 20;282(16):11795-804
CHIP (carboxyl terminus of Hsc70-interacting protein) promotes basal and geldanamycin-induced degradation of estrogen receptor-alpha.
Fan M, Park A, Nephew KP
Molecular endocrinology (Baltimore, Md.) 2005 Dec;19(12):2901-14
Molecular endocrinology (Baltimore, Md.) 2005 Dec;19(12):2901-14
CHIP-Hsc70 complex ubiquitinates phosphorylated tau and enhances cell survival.
Shimura H, Schwartz D, Gygi SP, Kosik KS
The Journal of biological chemistry 2004 Feb 6;279(6):4869-76
The Journal of biological chemistry 2004 Feb 6;279(6):4869-76
Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions.
Ballinger CA, Connell P, Wu Y, Hu Z, Thompson LJ, Yin LY, Patterson C
Molecular and cellular biology 1999 Jun;19(6):4535-45
Molecular and cellular biology 1999 Jun;19(6):4535-45
Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions.
Ballinger CA, Connell P, Wu Y, Hu Z, Thompson LJ, Yin LY, Patterson C
Molecular and cellular biology 1999 Jun;19(6):4535-45
Molecular and cellular biology 1999 Jun;19(6):4535-45
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CHIP was performed by loading 40 µg of MCF-7 cell lysates and 3 µL PageRuler Prestained Protein Ladder (Product # 26619) per well onto a 10% Tris-HCl polyacrylamide gel. Proteins were transferred to a NC membrane and blocked with 5% skim milk/PBS-T for 1 h at room temperature. CHIP was detected at 35 kDa using a CHIP antibody (Product # PA1-015) at 1:1000 dilution in 5% BSA/PBS-T buffer overnight at 4°C on a rocking platform, followed by a goat anti-rabbit IgG-HRP secondary antibody at a dilution in 5% skim milk/PBS-T of 1:3000 for 2 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). Data courtesy of Dr. Wei Xu at the University of Wisconsin, Madison, WI.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot detection of transfected COS-1 cells expressing CHIP using Product # PA1-015.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Carboxyl Terminus of Hsc70-Interacting Protein (CHIP) was performed by loading 20 µg of total protein extracted from the left hemisphere (Left lane)or right hemisphere (right lane) of a normal C57BL/6 mouse brain per well on an SDS-PAGE gel. Proteins were transferred to a membrane, blocked with 5% non-fat dry milk, and probed with a CHIP polyclonal antibody (Product # PA1-015) at a dilution of 1:1000, followed by a HRP-conjugated goat anti-rabbit IgG secondary antibody. Detection was performed using a chemiluminescent substrate. Data courtesy of the Innovators Program.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CHIP was performed by loading 40 µg of MCF-7 cell lysates and 3 µL PageRuler Prestained Protein Ladder (Product # 26619) per well onto a 10% Tris-HCl polyacrylamide gel. Proteins were transferred to a NC membrane and blocked with 5% skim milk/PBS-T for 1 h at room temperature. CHIP was detected at 35 kDa using a CHIP antibody (Product # PA1-015) at 1:1000 dilution in 5% BSA/PBS-T buffer overnight at 4°C on a rocking platform, followed by a goat anti-rabbit IgG-HRP secondary antibody at a dilution in 5% skim milk/PBS-T of 1:3000 for 2 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). Data courtesy of Dr. Wei Xu at the University of Wisconsin, Madison, WI.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
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- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 Searching for the E3-ligase that mediates degradation of BRCA2 upon hyperthermia. ( A ) Effect of hyperthermia on BRCA2 levels upon knock-down of the Heat-shock Protein (HSP)-associated E3-ligase, C terminus of H SC70- I nteracting P rotein (CHIP), in U2OS cells using two different siRNAs. Arrow on the right next to the CHIP panel indicates the predicted position of the CHIP protein. ( B ) Effect of hyperthermia on BRCA2 levels upon knock-down of the SUMO-E3 ligases Protein Inhibitor Of Activated STAT( PIAS) 1-4 in U2OS cells using siRNAs. ( C ) Effect of hyperthermia on BRCA2 levels upon siRNA knock-down of DNA-repair-related, SUMO-targeted ubiquitin ligases, Ring Finger (RNF) 4 and RNF111. ( D ) Detection of BRCA2 and BRCA1 in UWB1.289 (BRCA1-) cells with or without complementation of wild type BRCA1 (wtBRCA1). ( E ) Detection of BRCA2 in U2OS cells treated with increased doses of the neddylation-inhibitor MLN4924, added 30 min prior to hyperthermia. ( F ) Detection of BRCA2 in U2OS cells treated with or without hyperthermia in the presence of proteasome inhibitor MG132 (10 uM) or the E1-activating enzyme inhibitor PYR-41 (10 uM). Inhibitors were added 1 h prior to hyperthermia treatment, and left on for the duration of the treatment.
