Qty: 100 µg/200 µL
Mouse anti-BAG-1
Catalog No. 37-4000
Lot No.
Mouse anti-BAG-1
FORM
This monoclonal antibody is supplied as a 200 µL aliquot at a concentration of 0.5 mg/mL in PBS, pH 7.4, containing 0.1%
sodium azide. This antibody is highly purified from mouse ascites by protein A affinity.
ISOTYPE: Mouse IgG1-kappa
CLONE: 10B6H7
IMMUNOGEN
Recombinant human GST-BAG-1
SPECIFICITY
This antibody is specific for the human BAG-1, BAG-1M and BAG-1L proteins. On Western blots, it identifies the target band
at ~35 kDa.
REACTIVITY
Reactivity has been confirmed with human LNCaP, PC3, and Jurkat cells, paraffin-embedded human breast cancer tissues,
as well as mouse heart, lung and spinal cord homogenates and WEH/231, Ag8 and 32D.3 cell lysates.
Sample
ELISA
Human
Mouse
Immunogen
ND
ND
+++
Immunoprecipitation
(native)
++
++
ND
Immunofluorescence
Western
Blotting
++
++
ND
+++
+++
ND
Immuno
histochemistry
(frozen)
++
++
ND
Immuno
histochemistry
(paraffin)*
++
++
ND
(Excellent +++, Good++, Poor +, No reactivity 0, Not applicable N/A, Not Determined ND)
USAGE
Working concentrations for specific applications should be determined by the investigator. Appropriate concentrations will be
affected by several factors, including secondary antibody affinity, antigen concentration, sensitivity of detection method,
temperature and length of incubations, etc. The suitability of this antibody for applications other than those listed below has
not been determined. The following concentration ranges are recommended starting points for this product.
ELISA:
Immunoprecipitation:
Western Blotting:
Immunofluorescence:
Immunohistochemistry*:
0.1 – 1.0 ug/mL
10 µg/test
1-5 µg/mL
1-5 µg/mL
5-10 µg/mL
*For immunohistochemistry with paraffin-embedded tissues, heat induced epitope retrieval (HIER) with citrate buffer, pH 6.0, is required.
For IHC in formalin-fixed, paraffin-embedded tissues, Cat. No. 18-7481 is recommended.
STORAGE
Store at 2-8°C for up to one month. Store at –20°C for long-term storage. Avoid repeated freezing and thawing.
(cont’d)
www.invitrogen.com
Invitrogen Corporation • 542 Flynn Rd • Camarillo • CA 93012 • Tel: 800.955.6288 • E-mail: techsupport@invitrogen.com
PI374000
(Rev 10/08) DCC-08-1089
Important Licensing Information - These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen Catalog or our website,
www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these
products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
(37-4000 cont’d)
BACKGROUND
BAG-1 is a multifunctional protein that blocks apoptosis and interacts with several types of proteins, including Bcl-2 family
proteins, the kinase Raf-1, certain tyrosine kinase growth factor receptors, and steroid hormone receptors, possibly by virtue
1,2
of its ability to regulate the Hsp70/Hsc70 family of molecular chaperones . Two major forms of the human and mouse BAG1 proteins have been detected by immunoblotting. The longer human and mouse BAG-1 proteins (BAG-1L) appear to arise
through translation initiation at noncanonical CTG codons located upstream of and in-frame with the usual ATG codon used
for production of the originally described BAG-1 protein. In normal tissues, BAG-1L is far more restricted in its expression
and is present at lower levels than the more prevalent BAG-1 protein. Human but not mouse tissues also produce small
amounts of an additional isoform of BAG-1 of intermediate size (BAG-1M) that probably arises through translation initiation at
yet another site involving an ATG codon. All three isoforms of human BAG-1 (BAG-1, BAG-1M, and BAG-1L) retain the
ability to bind Hsc70. Subcellular fractionation and immunofluorescence confocal microscopy studies have indicated that
BAG-1L often resides in the nucleus, consistent with the presence of a nuclear localization sequence in the N-terminal
unique domain of this protein. In immunohistochemical assays, BAG-1 reactivity was detected in a wide variety of cell types
in normal adult tissues and was localized to cytosol, nucleus, or both, depending on the particular type of cell.3 In some
cases, cytosolic BAG-1 immunostaining was clearly associated with organelles resembling mitochondria, consistent with the
3
reported interaction of BAG-1 with Bcl-2 and related proteins.
The BAG-1 protein was found at high levels in several types of human tumor cell lines among the 67 tested, particularly
leukemias, breast, prostate, and colon cancers.3 In contrast to normal tissues, which only rarely expressed BAG-1L, tumor
cell lines commonly contained BAG-1L protein, including most prostate, breast, and leukemia cell lines, suggesting that a
change in BAG-1 mRNA translation frequently accompanies malignant transformation.3
REFERENCES
1.
2.
3.
Stuart JK, et al. J Biol Chem 273(35):22506-22514, 1998.
Bimston D, et al. EMBO J 17(23):6871-6878, 1998.
Takayama S, et al. Cancer Res 58:3116-3131, 1998.
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For Research Use Only
TM022604
www.invitrogen.com
Invitrogen Corporation • 542 Flynn Rd • Camarillo • CA 93012 • Tel: 800.955.6288 • E-mail: techsupport@invitrogen.com
PI374000
(Rev 10/08) DCC-08-1089
Important Licensing Information - These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen Catalog or our website,
www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these
products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.