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rabbit anti-PUMA (CT) polyclonal antibody 7882

$445.00

Antibody summary

  • Rabbit polyclonal to PUMA (CT)
  • Suitable for: ELISA,WB,ICC,IF
  • Isotype: IgG
  • 100 µg
SKU: 7882parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

PUMA (CT)

available sizes

100 µg

Available product – rabbit anti-PUMA (CT) polyclonal antibody 7882

antibody
Tested applications
WB,ICC/IF,ELISA
Recommended dilutions
Immunoblotting: use at 2ug/mL.

Positive control: K562 cell lysate.

Immunocytochemistry: use at 1ug/mL.

These are recommended concentrations.

Enduser should determine optimal concentrations for their applications.
Immunogen
Synthetic peptide corresponding to aa 180-193 of human PUMA-a (accession no NP_055232). This sequence is identical in human PUMA-a and PUMA-b.
Size and concentration
100µg and lot specific
Form
liquid
Storage Instructions
This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Storage buffer
PBS, pH 7.4.
Purity
peptide affinty purifcation
Clonality
polyclonal
Isotype
IgG
Compatible secondaries
goat anti-rabbit IgG, H&L chain specific, peroxidase conjugated, conjugated polyclonal antibody 9512
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody 2079
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody 7863
goat anti-rabbit IgG, H&L chain specific, Cross Absorbed polyclonal antibody 2371
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1715
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1720
Isotype control
Rabbit polyclonal - Isotype Control
target relevance
Protein names
Bcl-2-binding component 3, isoforms 1/2 (JFY-1) (p53 up-regulated modulator of apoptosis)
Gene names
BBC3,BBC3 PUMA
Protein family
Bcl-2 family
Mass
20532Da
Function
FUNCTION: Essential mediator of p53/TP53-dependent and p53/TP53-independent apoptosis (PubMed:11463391, PubMed:23340338). Promotes partial unfolding of BCL2L1 and dissociation of BCL2L1 from p53/TP53, releasing the bound p53/TP53 to induce apoptosis (PubMed:23340338). Regulates ER stress-induced neuronal apoptosis (By similarity). {ECO:0000250|UniProtKB:Q99ML1, ECO:0000269|PubMed:11463391, ECO:0000269|PubMed:23340338}.
Subellular location
SUBCELLULAR LOCATION: Mitochondrion {ECO:0000269|PubMed:11463391, ECO:0000269|PubMed:11463392, ECO:0000269|PubMed:11572983}. Note=Localized to the mitochondria in order to induce cytochrome c release.
Tissues
TISSUE SPECIFICITY: Ubiquitously expressed. {ECO:0000269|PubMed:11463391}.
Structure
SUBUNIT: Interacts with MCL1 and BCL2A1 (By similarity). Interacts (via BH3 domain) with BCL2 (PubMed:11463391). Interacts with BCL2L1/BCL-XL (PubMed:23340338). Interacts (via BH3 domain) with NOL3/ARC (via CARD domain); this interaction prevents BBC3 association with BCL2 and results in CASP8 activation (By similarity). {ECO:0000250|UniProtKB:Q80ZG6, ECO:0000250|UniProtKB:Q99ML1, ECO:0000269|PubMed:11463391, ECO:0000269|PubMed:23340338}.
Target Relevance information above includes information from UniProt accession : Q9BXH1
The UniProt Consortium

Data

benchmark-antibodies_anti-puma_ct_antibody_7882_1.gif
Western Blot Validation of PUMA in K562 and 3T3/NIH Cells
Loading: 15 µg of lysates per lane. Antibodies: 7882 (2 µg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-puma_ct_antibody_7882_2.gif
Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 20 µg of lysates per lane. Antibodies: 7882 (3 µg/mL), 11008 (2 µg/mL), beta-actin (1 µg/mL) and GAPDH (0.02 µg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-puma_ct_antibody_7882_3.gif
Validation with PUMA siRNA Knockdown in 293 Cells
293 cells were transfected with control siRNAs (lane 1) or PUMA siRNAs (lane 2) Loading: 15 µg of 293 whole cell lysates per lane. Antibodies: 7882 (2 µg/mL), beta-actin (1 µg/mL) and GAPDH (0.02 µg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-puma_ct_antibody_7882_4.gif
Sensitivity Test for PUMA in 2983 Cells
Loading: Lysates/proteins at 15 µg per lane. Antibodies: 7882 (lane 1-3: 1, 2 and 4 µg/mL). 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-puma_ct_antibody_7882_5.jpg
Immunofluorescence Validation of PUMA in K562 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling PUMA with 7882 at 2 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing cytosol staining on K562 cells.
benchmark-antibodies_anti-puma_ct_antibody_7882_6.jpg
Immunocytochemistry Validation of PUMA in K562 Cells
Immunocytochemical analysis of K562 cells using anti-PUMA antibody (7882) at 1 µg/mL. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-puma_ct_antibody_7882_7.gif
KD Validation of PUMA in HOSE-RasV12 Cells (Elgendy et al., 2011)
HOSE-RasV12 cells were transfected with control shRNA plasmid or shRNA plasmids (KD) targeted against Noxa or Puma, as indicated. PUMA expression was not observed in PUMA KD cells detected by anti-PUMA antibodies (7882).
benchmark-antibodies_anti-puma_ct_antibody_7882_8.gif
Induction Validation of PUMA in Primary Cortical Neurons (Sabirzhanov et al., 2014)
PUMA protein levels were increased in etoposide-treated primary cortical neurons detected by anti-PUMA antibodies (7882).
benchmark-antibodies_anti-puma_ct_antibody_7882_9.gif
KD Validation of PUMA PUMA in Tet Cells (Han et al., 2010)
Immunoblot analyses of Tet-induced p53 cells treated with NOXA, Puma, Bim or non-targeting siRNAs that were utilized in this experiment. PUMA protein levels were markedly reduced in PUMA KD cells detected by anti-PUMA antibodies (7882).

Publications

Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.




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Protocols

relevant to this product
Western blot
IHC
ICC

Documents

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