Western Blot: ID4 Antibody [H00003400-B01P] - Analysis of ID4 expression in transfected 293T cell line by ID4 polyclonal antibody. Lane 1: ID4 transfected lysate(17.71 KDa). Lane 2: Non-transfected lysate.
Antibody reactivity against Recombinant Protein with GST tag on ELISA and WB and also on transfected lysate in WB. GST tag alone is used as a negative control. ICC/IF usage reported in scientific literature (PMID: 25778840).
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Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
PBS (pH 7.4)
Protein A purified
This product is produced by and distributed for Abnova, a company based in Taiwan.
Alternate Names for ID4 Antibody
Class B basic helix-loop-helix protein 27
DNA-binding protein inhibitor ID-4
Inhibitor of DNA binding 4
inhibitor of DNA binding 4, dominant negative helix-loop-helix protein
Transcription factors containing a basic helix-loop-helix (bHLH) motif regulate expression of tissue-specific genes in a number of mammalian and insect systems. DNA-binding activity of the bHLH proteins is dependent on formation of homo- and/or heterodimers. Dominant-negative HLH proteins encoded by Id-related genes, such as ID4, also contain the HLH-dimerization domain but lack the DNA-binding basic domain. Consequently, Id proteins inhibit binding to DNA and transcriptional transactivation by heterodimerization with bHLH proteins (Pagliuca et al., 1995 [PubMed 7665172]).[supplied by OMIM]
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.
Tested in gray short-tailed opossum (Monodelphis domestica) in testis from 8-month animal. Tissue was fixed in 4% Paraformaldehyde overnight, incubated in 30% sucrose/PBS overnight and embedded in OCT. Sections were cut on a cryostat at 10um.
For immunofluorescence staining: Slides were dried 15 minutes, washed 3x3 min TBST (0.1% Triton X-100), permeabilized for 10 minutes in 2% Triton X-100 in TBS, rinsed 3x3 min TBST, blocked in 10% heat inactivated goat serum in TBST for 30 min, and primary added at 1:100 in 10% blocking solution. Slides were coverslipped with parafilm overnight at 4C. The next day slides were washed with TBST 4x5 min, secondaries (AlexaFluor 568 goat anti rabbit and AlexaFluor 488 goat anti mouse 1:500) for 1 hour, 3x3 washes TBST, and mounted with VectaShield HardSet (+ DAPI).
Intense staining at periphery of seminiferous tubules of Uchl-1 (red in image) with these conditions. 1:200 also worked though staining was less intense (not shown). The same conditions with 1% Triton X-100 for permeabiliziation resulted in weak staining at 1:100 and 1:200. However, ID-4 did not work well under any conditions. Shown in green is some faint staining in interstitial cells but this was not consistent between samples and did not appear to be specific. ID-4 was tested in 4 and 8 month opossum testes, and at dilutions ranging from 1:25 to 1:500. Single-antibody staining was also performed for each antibody and successful in Uchl-1, not ID-4.
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