Endophilin B1/Bif-1 Antibody (30A882.1.1) - BSA Free

Western Blot: Endophilin B1/Bif-1 Antibody (30A882.1.1) [NBP2-24733] - Amino acid E28 on Bif-1 is important for calpain-mediated processing and autophagic clearance of p62. The myc-tagged constructs: wild-type Endophilin B1/Bif-1 and E28A Endophilin B1/Bif-1 were transfected into H1299 cells. 24h later, the cells were treated or not with 100nM thapsigargin for 1h. Afterwards, the cell lysates were prepared and analyzed by western blot with the indicated antibodies. Image collected and cropped by CiteAb from the following publication (//bio.biologists.org/content/6/5/551.long), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: Endophilin B1/Bif-1 Antibody (30A882.1.1) [NBP2-24733] - IHC analysis of formalin fixed paraffin-embedded (FFPE) human kidney using 5 ug/ml conc. of Endophilin B1 antibody (clone 30A882.1.1) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Cytoplasmic staining of Endophilin B1 was observed. Staining was performed by Histowiz.

Western Blot: Endophilin B1/Bif-1 Antibody (30A882.1.1) [NBP2-24733] - Analysis for Bi-f-1/Endophilin-B1 using this antibody at 2 ug/ml dilution against 10 ug of Jurkat cell lysate.

Simple Western: Endophilin B1/Bif-1 Antibody (30A882.1.1) [NBP2-24733] - Simple Western lane view shows a specific band for Endophilin B1/Bif-1 in 0.05 mg/ml of Jurkat lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF, IHC, IP
• Clone: 30A882.1.1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Endophilin B1/Bif-1 Antibody (30A882.1.1) - BSA Free Summary

• Immunogen: A full-length human Bif-1 recombinant protein was used as immunogen.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: SH3GLB1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence reported in multiple pieces of scientific literature
  • Immunohistochemistry 5ug/ml
  • Immunohistochemistry-Paraffin 5 ug/ml
  • Immunoprecipitation 1:10-1:500
  • Simple Western 1:200
  • Western Blot
• Application Notes: In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in Jurkat lysate 0.05 mg/mL, separated by Size, antibody dilution of 1:200, apparent MW was 47 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reactivity Notes

Use in Human reported in scientific literature (PMID:33802405).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Endophilin B1/Bif-1 Antibody (30A882.1.1) - BSA Free

• Bax-interacting factor 1
• Bif1
• Bif-1
• Bif-1dJ612B15.2
• CGI-61
• Endophilin B1
• endophilin-B1
• KIAA0491endophilin B1
• SH3 domain-containing GRB2-like protein B1
• SH3-containing protein SH3GLB1
• SH3-domain GRB2-like endophilin B1
• SH3-domain, GRB2-like, endophilin B1
• SH3GLB1

Background

Bif-1/Endophilin-B1 may be required for normal outer mitochondrial membrane dynamics. Required for coatomer-mediated retrograde transport in certain cells. May recruit other proteins to membraneswith high curvature. May promote membrane fusion

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Endophilin B1/Bif-1 Antibody (NBP2-24733)(31)

Publications using NBP2-24733

• Hou P, Guo Y, Jin H et al. Bif-1c Attenuates Viral Proliferation by Regulating Autophagic Flux Blockade Induced by the Rabies Virus CVS-11 Strain in N2a Cells Microbiology spectrum 2023-04-04 [PMID: 37014208] (WB, Mouse)
• Cho C, Zeigler M, Mizuno S et al. Reductions in Hydrogen Sulfide and Changes in Mitochondrial Quality Control Proteins Are Evident in the Early Phases of the Corneally Kindled Mouse Model of Epilepsy International journal of molecular sciences 2022-01-27 [PMID: 35163358] (WB, Mouse)
• Cho C, Zeigler M, Mizuno S Et al. Reductions in Hydrogen Sulfide Precede Onset of Mitochondrial Quality Control in the Corneal Kindled Mouse Model of Epilepsy Preprints 2021-10-28 [PMID: 35163358] (WB, Mouse)
• Frankowski H, Yeboah F, Berry B et al. Knock-Down of HDAC2 in Human Induced Pluripotent Stem Cell Derived Neurons Improves Neuronal Mitochondrial Dynamics, Neuronal Maturation and Reduces Amyloid Beta Peptides International Journal of Molecular Sciences 2021-03-03 [PMID: 33802405] (WB, Human)
• Frangez , Seyed Jafari S, Hunger R, Simon H Loss of Concurrent Regulation of the Expression of BIF-1, BAX, and Beclin-1 in Primary and Metastatic Melanoma Biochemistry Moscow 2020-10-01 [PMID: 33202207]
• Cho SG, Xiao X, Wang S et al Bif-1 Interacts with Prohibitin-2 to Regulate Mitochondrial Inner Membrane during Cell Stress and Apoptosis. J Am Soc Nephrol. 2019-07-01 [PMID: 31126972] (WB, IP, Human)
  
  Details:
  Citation using the Azide Free version of this antibody.
• Riera, AM;Martinez, CI;Itskanov, S;Steffen, J;Roach, B; Dynamin-2 controls phagophore maturation bioRxiv 2019-09-02 (WB, ICC/IF)
• FrangeZ Z, FernAndez-Marrero Y, Stojkov D et al. BIF-1 inhibits both mitochondrial and glycolytic ATP production: its downregulation promotes melanoma growth Oncogene 2020-06-01 [PMID: 32493957] (IHC-P, Human)
• Frankowski H, Yeboah F, Berry B, et al. Histone Deacetylase 2 (HDAC2) influences maturation and mitochondrial dynamics in human induced pluripotent stem cell derived neurons bioRxiv 2021-01-19
• Raimondi M, Cesselli D, Di Loreto C et al. USP1 (ubiquitin specific peptidase 1) targets ULk1 and regulates its cellular compartmentalization and autophagy. Autophagy. 2018-10-18 [PMID: 30335599] (ICC/IF, Human)
• Wang DB, kinoshita C, kinoshita Y et al. Neuronal Susceptibility to Beta-Amyloid Toxicity and Ischemic Injury Involves Histone Deacetylase-2 Regulation of Endophilin-B1. Brain Pathol. 2018-07-20 [PMID: 30028551] (WB, Mouse)
• Serfass JM, Takahashi Y, Zhou Z et al. Endophilin B2 facilitates endosome maturation in response to growth factor stimulation, autophagy induction, and influenza A virus infection J. Biol. Chem. 2017-06-16 [PMID: 28455444] (ICC/IF, WB, Human)
• Marcassa E, Raimondi M, Anwar T et al. Calpain mobilizes Atg9/Bif-1 vesicles from Golgi stacks upon autophagy induction by thapsigargin. Biol Open. 2017-03-16 [PMID: 28302665] (WB, Human)
• Zhang C, Liu F, Chen H et al. Bif-1 promotes tumor cell migration and metastasis via Cdc42 expression and activity. Clin. Exp. Metastasis 2016-10-11 [PMID: 27730394] (IF/IHC, Human)
• Wild F, Khan MM, Straka T, Rudolf R. Progress of endocytic CHRN to autophagic degradation is regulated by RAB5-GTPase and T145 phosphorylation of SH3GLB1 at mouse neuromuscular junctions in vivo. Autophagy 2016-10-07 [PMID: 27715385] (WB, Mouse)
• Wang YH, Wang JQ, Wang Q et al. Endophilin B2 promotes inner mitochondrial membrane degradation by forming heterodimers with Endophilin B1 during mitophagy. Sci Rep. 2016-04-26 [PMID: 27112121]
• Takahashi Y, Tsotakos N, Liu Y et al. The Bif-1-Dynamin 2 membrane fission machinery regulates Atg9-containing vesicle generation at the Rab11-positive reservoirs. Oncotarget. 2016-04-12 [PMID: 26980706] (WB, IP, Human)
• Xu S, Cherok E, Das S et al. Mitochondrial E3 ubiquitin ligase MARCH5 controls mitochondrial fission and cell sensitivity to stress-induced apoptosis through regulation of MiD49 protein. Mol. Biol. Cell. 2015-11-12 [PMID: 26564796] (WB, Human)
• Wang DB, Kinoshita Y, Kinoshita C et al. Loss of endophilin-B1 exacerbates Alzheimer's disease pathology. Brain. 2015-05-16 [PMID: 25981964] (WB, Mouse, Human)
  
  Details:
  Endophilin B1/Bif-1 antibody (clone 30A882.1.1) used for WB application at 1:500 dilution in experiments involving a mouse model of Alzheimer’s disease pathology. Figure 1 shows the expression of Endophilin B1/Bif-1 in lysates from hippocampus and cortex of 10-month-old double mutant amyloid precursor protein and presenilin 1 (APPswe/PSEN1dE9) mouse model of Alzheimer’s disease and wild-type littermates. WB also performed on postnatal cortical neurons which were infected with EGFP or endophilin-B1 isoforms for 3 days, then treated with 10 mM amyloid-b25-35 or reverse peptide amyloid-b35-25 for 24 h (Figure 6A). Expression of Endophilin B1 was also studied in the inferior parietal lobule cerebral cortex of patients at various Braak stages (Figure 7). In all WB results, this antibody generated two major bands with the lower band representing ubiquitously expressed endophilin-B1a isoform and the upper band corresponding to neuron-specific isoforms, endophilin-B1b/c (described previously also by Wang et al., 2014; PMID 24523556).
• Dong H, Tian L, Li R et al. IFNg-induced Irgm1 promotes tumorigenesis of melanoma via dual regulation of apoptosis and Bif-1-dependent autophagy Oncogene. 2015-01-26 [PMID: 25619828] (WB, Mouse)
  
  Details:
  Bif-1/Endophilin-B1 / SH3GLB1 antibody used for WB assay on lysates of Irgm1-overexpressing B16 cells that were transfected with control siRNA or Bif-1-siRNA followed by 6 or 24 hours of starvation with serum-deprived low-glucose conditioned medium treatment (Figure 7a).
• Wang DB, Uo T, Kinoshita C et al. Bax interacting factor-1 promotes survival and mitochondrial elongation in neurons. J. Neurosci. 2014-02-12 [PMID: 24523556] (WB, ICC/IF, Mouse)
  
  Details:
  Primary mouse neurons and astrocytes, mouse embryo fibroblasts, 3T3 cells. WB: Figs 1, 4b, 5a,d,f,6e, 7. IF: Fig 6a. Notes: 1. The Bif-1 mAb is a non-isoform-specific antibody and recognized all isoforms evaluated. 2. The specificity of the antibody was v
• Runkle Kristin B, Meyerkord Cheryl L, Desai Neelam V et al. Bif-1 suppresses breast cancer cell migration by promoting EGFR endocytic degradation. Cancer Biol Ther. 2012-08-01 [PMID: 22785202] (Human)
• Cant JS, Goodale MA. Scratching beneath the surface: new insights into the functional properties of the lateral occipital area and parahippocampal place area. J Neurosci 2011-06-01 [PMID: 21632946] (ICC/IF, Human)
• Takahashi Y, Meyerkord CL, Hori T et al. Bif-1 regulates Atg9 trafficking by mediating the fission of Golgi membranes during autophagy. Autophagy 2011-01-01 [PMID: 21068542] (ICC/IF)
• Yang J, Takahashi Y, Cheng E et al. GSK-3beta promotes cell survival by modulating Bif-1-dependent autophagy and cell death. J Cell Sci. 2010-03-15 [PMID: 20159967] (WB)
  
  Details:
  Bif-1 (IMG-265A) WB, Fig 5A, 5B, 5C, 5D, PC-3 cells. IP, Fig 6D, PC-3 cells.
• Etxebarria A, Terrones O, Yamaguchi H et al. Endophilin B1/Bif-1 stimulates BAX activation independently from its capacity to produce large scale membrane morphological rearrangements. J Biol Chem. 2009-02-13 [PMID: 19074440] (WB, Rat)
  
  Details:
  WB (rat liver mitochondria), Fig. 1A; 2A,B,C; 3D; 4B.
• Yamaguchi H, Woods NT, Dorsey JF et al. SRC directly phosphorylates Bif-1 and prevents its interaction with Bax and the initiation of anoikis. J Biol Chem. 2008-07-04 [PMID: 18474606] (IP, Human)
  
  Details:
  IP (human 293T, A431, and NIH3T3 cell lines), Fig. 1C; 4D; 5C,D.
• Takahashi Y, Coppola D, Matsushita N et al. Bif-1 interacts with Beclin 1 through UVRAG and regulates autophagy and tumorigenesis. Nat Cell Biol. 2007-10-01 [PMID: 17891140]
• Takahashi Y, Karbowski M, Yamaguchi H et al. Loss of Bif-1 suppresses Bax/Bak conformational change and mitochondrial apoptosis. Mol Cell Biol 2005-11-01 [PMID: 16227588]
• Karbowski M, Jeong SY, Youle RJ. Endophilin B1 is required for the maintenance of mitochondrial morphology. J Cell Biol. 2004-09-27 [PMID: 15452144]
• Cuddeback SM, Yamaguchi H, Komatsu K et al. Molecular cloning and characterization of Bif-1. A novel Src homology 3 domain-containing protein that associates with Bax. J Biol Chem. 2001-06-08 [PMID: 11259440]

Bioinformatics

• Gene Symbol: SH3GLB1
• Uniprot:
  • Q9Y371()